Solid oral formulation of fenretinide

ABSTRACT

Amorphous solid dispersions suitable for oral delivery comprising fenretinide or an analog thereof and at least one matrix polymer, and processes for making the dispersions, are disclosed. Also disclosed are solid oral formulations comprising the amorphous solid dispersions, as well as uses thereof for the prevention and/or treatment of diseases or conditions treatable by fenretinide, including but not limited to cancers, conditions associated with a lipid imbalance, cystic fibrosis, osteoporosis, conditions associated with inflammation or opportunistic infections, and other diseases such as diabetes, obesity, dry-form age-related macular degeneration.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a Divisional Application of U.S. patent applicationSer. No. 15/328,381, filed on Jan. 23, 2017, which is a National EntryApplication of PCT application no. PCT/CA2015/000445 filed on Jul. 24,2015 and published in English under PCT Article 21(2), which itselfclaims the benefit of U.S. provisional application Ser. No. 62/029,119,filed on Jul. 25, 2014, each of which are incorporated by reference intheir entireties.

TECHNICAL FIELD

The present invention generally relates to pharmaceutical formulations,and more specifically to oral pharmaceutical compositions comprising theactive ingredient fenretinide or analogs thereof.

BACKGROUND ART

Fenretinide (all-trans-N-(4-hydroxyphenyl) retinamide; also referred toas 4-HPR), which has CAS registry number 65646-68-6, is a syntheticretinoid. Fenretinide was initially developed as a less toxic and bettertolerated derivative of retinoic acid and has been extensively studiedbecause of its chemo-protective and anti-tumor activities described whenused on a variety of malignant cells, including non-small cell lungcancer, neuroblastoma, Kaposi's sarcoma, breast cancer and glioma(Charles, et al. (2001) Cancer Chemother. Pharmacol. 47:444-450;Garaventa, et al. (2003) Clin. Cancer Res. 9:2032-2039; Lippman, et al.(2001) J. Natl. Cancer Inst. 93:605-618; Ponthan, et al. (2003) Oncol.Rep. 10:1587-1592; Puduvalli, et al. (1999) Clin. Cancer Res.5:2230-2235; Rao, et al. (1998) Breast Cancer Res. Treat. 48:265-271),and has been approved for clinical trials of cancer patients. However,despite its promising anticancer activity in preclinical studies, itslimited oral bioavailability, notably due to its poor water solubility,represents a significant challenge for its clinical assessment.

Fenretinide has been formulated in corn oil-containing soft-gelatincapsules, but such formulations have been shown to result in variableand low systemic exposures (i.e. poor bioavailability). Also, because oftheir size, patient compliance has been shown to be a concern with thesecorn oil-containing capsules, especially in pediatric subjects.Fenretinide has also been formulated in a lipid matrix, Lym-X-Sorb(LXS), (Maurer B J, Clin Cancer Res 13: 3079-3086, 2007), administratedas an oral powder delivered in non-milk fat-containing foods, andespecially as a slurry in non-milk fat-containing, or soy-basednutritional supplements. However, this formulation has been shown to beassociated with significant gastrointestinal (GI) side-effects,especially at higher doses (Kummar et al. (2011) Anticancer Research31(3):961-966), as well as to significant patient withdrawal due to thetaste and texture of the medication.

There is thus a need for new pharmaceutical compositions of fenretinide,especially for oral administration, capable to overcome the poor oralbioavailability of corn-oil based formulation, while allowing for morecompliant pharmaceutical dosage forms such as hard gelatine capsules,tablets, strips, caplets, suspensions, or powders for suspensions.

The present description refers to a number of documents, the content ofwhich is herein incorporated by reference in their entirety.

SUMMARY OF THE INVENTION

In a first aspect, the present invention relates to the preparation oforal formulations which comprises the amorphous state of the activeingredient fenretinide or analogs thereof. Such chemically and/orphysically stable formulations are composed of solid dispersions (e.g.,spray-dried), such as microparticles which contain fenretinide or one ofits analogs, where its amorphous state is maintained over time. Itsamorphous state is associated with improved oral bioavailabilityfollowing oral administration, when compared to crystalline fenretinide.The amorphous solid dispersion of fenretinide can beorally-administrated in an acceptable pharmaceutical form such as hardgelatine capsule, tablets, strips, caplets, cachets, lozenges,suspensions, or powders for suspensions.

In other aspects, the present invention relates to the following items 1to 116:

1. An amorphous solid dispersion for oral delivery comprisingfenretinide or an analog thereof and at least one matrix polymer.

2. The amorphous solid dispersion of item 1, wherein the at least onematrix polymer is a polyvinylpyrrolidone, a hydroxypropyl cellulose, ahydroxypropyl methylcellulose hypromellose phthalate, apolyvinylpyrrolidone-vinyl acetate, a hypromellose-acetate-succinate, orany mixture thereof.

3. The amorphous solid dispersion of item 2, wherein thepolyvinylpyrrolidone polymer is polyvinylpyrrolidone K12,polyvinylpyrrolidone K17, polyvinylpyrrolidone K25, polyvinylpyrrolidoneK30, polyvinylpyrrolidone K90, or any mixture thereof.

4. The amorphous solid dispersion of any one of items 1 to 3, whereinthe fenretinide or analog thereof is present in an amount in the rangeof about 20% to about 60% by weight.

5. The amorphous solid dispersion of item 4, wherein the fenretinide oranalog thereof is present in an amount in the range of about 30% toabout 50% by weight.

6. The amorphous solid dispersion of item 5, wherein the fenretinide oranalog thereof is present in an amount of about 40% by weight.

7. The amorphous solid dispersion of any one of items 1 to 6, where theamorphous state is obtained by fast evaporation, spray-drying,precipitation or melt extrusion.

8. The amorphous solid dispersion of item 7, where the amorphous stateis obtained by spray-drying

9. The amorphous solid dispersion of any one of items 1 to 8, comprisingfenretinide.

10. The amorphous solid dispersion of any one of items 1 to 9, furthercomprising an antioxidant.

11. The amorphous solid dispersion of item 10, wherein the antioxidantis butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT),citric acid, sodium metabisulfite, alpha-tocopherol and/or L-ascorbicacid.

12. The amorphous solid dispersion of item 11, wherein the antioxidantis L-ascorbic acid.

13. A process for making the amorphous solid dispersion according to anyone of items 1 to 12, the process comprising:

(a) forming a solution comprising the fenretinide or analog thereof, theat least one matrix polymer, and a solvent or solvent mixture in whichboth the fenretinide or analog thereof and the at least one matrixpolymer are soluble; and

(b) spray-drying the solution of step (a), thereby obtaining theamorphous solid dispersion.

14. The process of item 13, where the solvent or solvent mixturecomprises dichloromethane, methanol and/or ethanol.

15. The process of item 14, where the solvent is dichloromethane,methanol or ethanol.

16. The process of any one of items 13 to 15, where the solution furthercomprises an antioxidant.

17. The process of item 16, wherein the antioxidant is butylatedhydroxyanisole (BHA), butylated hydroxytoluene (BHT), citric acid,sodium metabisulfite, and/or L-ascorbic acid.

18. The process of item 17, wherein the antioxidant is L-ascorbic acid.

19. A spray-dried amorphous solid dispersion obtained by the process ofany one of items 13 to 18.

20. An oral dosage formulation comprising the amorphous solid dispersionof any one of items 1-12 and 19.

21. The oral dosage formulation of item 20, wherein the amorphous soliddispersion is present in an amount from about 20 to about 80% by weight.

22. The oral dosage formulation of item 21, wherein the amorphous soliddispersion is present in an amount from 30 to 60% by weight.

23. The oral dosage formulation of item 22, wherein the amorphous soliddispersion is present in an amount from about 45 to about 55% by weight.

24. The oral dosage formulation of item 23, wherein the amorphous soliddispersion is present in an amount from about 50% by weight.

25. The oral dosage formulation of any one of items 20-24, wherein thefenretinide or analog thereof is present in an amount from about 10 toabout 250 mg per dose unit.

26. The oral dosage formulation of item 25, wherein the fenretinide oranalog thereof is present in an amount from about 25 mg to about 200 mgper dose unit.

27. The oral dosage formulation of item 26, wherein the fenretinide oranalog thereof is present in an amount from about 50 mg to about 150 mgper dose unit.

28. The oral dosage formulation of item 27, wherein the fenretinide oranalog thereof is present in an amount of about 100 mg per dose unit.

29. The oral dosage formulation of any one of items 20-28, furthercomprising at least one additional pharmaceutical excipient.

30. The oral dosage formulation of item 29, wherein said at least oneadditional pharmaceutical excipient comprises a disintegrant.

31. The oral dosage formulation of item 30, wherein said disintegrant isa cross-linked sodium carboxymethylcellulose.

32. The oral dosage formulation of item 30 or 31, wherein saiddisintegrant is present in an amount from about 2 to about 10% byweight.

33. The oral dosage formulation of item 32, wherein said disintegrant ispresent in an amount from about 4 to about 6% by weight.

34. The oral dosage formulation of any one of items 29-32, wherein saidat least one additional pharmaceutical excipient comprises a lubricant.

35. The oral dosage formulation of item 34, wherein said lubricantcomprises magnesium stearate.

36. The oral dosage formulation of item 34 or 35, wherein said lubricantis present in an amount from about 0.5 to about 2% by weight.

37. The oral dosage formulation of item 36, wherein said lubricant ispresent in an amount of about 1% by weight.

38. The oral dosage formulation of any one of items 29-37, wherein saidat least one additional pharmaceutical excipient comprises a filler orbinder.

39. The oral dosage formulation of item 38, wherein said filler orbinder comprises microcrystalline cellulose.

40. The oral dosage formulation of item 38 or 39, wherein said filler orbinder comprises calcium hydrogen phosphate dihydrate.

41. The oral dosage formulation of any one of items 38 to 40, whereinsaid filler or binder is present in an amount from about 20% to about45% by weight.

42. The oral dosage formulation of item 41, wherein said filler orbinder is present in an amount from about 30% to about 40% by weight.

43. The oral dosage formulation of any one of items 29-42, wherein saidat least one additional pharmaceutical excipient comprises anantioxidant.

44. The amorphous solid dispersion of item 43, wherein the antioxidantis butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT),citric acid, sodium metabisulfite, alpha-tocopherol and/or L-ascorbicacid.

45. The amorphous solid dispersion of item 44, wherein the antioxidantis L-ascorbic acid.

46. The oral dosage formulation of any one of items 20 to 45, which isin an acceptable pharmaceutical form for oral administration.

47. The oral dosage formulation of item 46, wherein said acceptablepharmaceutical form is a hard gelatine capsule, a tablet, a strip, acaplet, a sachet, a lozenge, a suspension, or a powder for suspension.

48. A method for preventing, treating, and/or lessening the severity ofcancer in a subject, said method comprising administering to a subjectin need thereof an effective amount of the amorphous solid dispersionaccording to any one of items 1-12 and 19, or the oral dosageformulation according to any one of items 20-47.

49. The method of item 48, wherein said cancer is breast cancer, ovariancancer, prostate cancer, cervical cancer, lung cancer, renal cancer,bladder cancer, glioma, skin cancer, head and neck carcinoma,non-Hodgkin's lymphoma, neuroblastoma, Ewing's sarcoma or Kaposi'ssarcoma.

50. A method for preventing, treating, and/or lessening the severity of,a disease or condition associated with a lipid imbalance, said methodcomprising administering to a subject in need thereof an effectiveamount of the amorphous solid dispersion according to any one of items1-12 and 19, or the oral dosage formulation according to any one ofitems 20-47.

51. The method of item 50, wherein said disease or condition associatedwith lipid imbalance is Cystic Fibrosis.

52. The method of item 50, wherein said disease or condition associatedwith lipid imbalance is a respiratory disease, a neural injury, or aneurodegenerative disease.

53. A method for preventing, treating, and/or lessening the severity ofCystic Fibrosis or a condition associated with Cystic Fibrosis, saidmethod comprising administering to a subject in need thereof aneffective amount of the amorphous solid dispersion according to any oneof items 1-12 and 19, or the oral dosage formulation according to anyone of items 20-47.

54. A method for preventing, treating, and/or lessening the severity ofa bone disease, or a condition associated with a bone disease, saidmethod comprising administering to a subject in need thereof aneffective amount of the amorphous solid dispersion according to any oneof items 1-12 and 19, or the oral dosage formulation according to anyone of items 20-47.

55. The method of item 54, wherein said subject suffers from CysticFibrosis.

56. A method for preventing, treating, and/or lessening the severity ofa disease or condition associated with inflammation, said methodcomprising administering to a subject in need thereof an effectiveamount of the amorphous solid dispersion according to any one of items1-12 and 19, or the oral dosage formulation according to any one ofitems 20-47.

57. The method of item 56, wherein said disease or condition associatedwith inflammation is a respiratory disease, a neural injury, or aneurodegenerative disease.

58. The method of item 56, wherein said inflammation is lunginflammation, and the disease or condition associated with lunginflammation is Cystic Fibrosis.

59. The method of item 57, wherein said neural injury is Spinal CordInjury (SCI)

60. The method of item 57, wherein said neurodegenerative disease isAmyotrophic Lateral Sclerosis (ALS).

61. A method for preventing, treating, and/or lessening the severity ofan opportunistic infection or a condition associated with anopportunistic infection, said method comprising administering to asubject in need thereof an effective amount of the amorphous soliddispersion according to any one of items 1-12 and 19, or the oral dosageformulation according to any one of items 20-47.

62. The method of item 61, wherein said condition associated with anopportunistic infection is a respiratory disease.

63. The method of items 62, wherein said respiratory disease is CysticFibrosis.

64. A method for preventing, treating, and/or lessening the severity ofa disease in a subject, wherein the disease is allergic asthma, SpinalCord Injury, Amyotrophic Lateral Sclerosis, diabetes, obesity, maculardegeneration, AIDS, allergic encephalomyelitis, ichthyosis or a viralinfection caused by a flavivirus, said method comprising administeringto a subject in need thereof an effective amount of the amorphous soliddispersion according to any one of items 1-12 and 19, or the oral dosageformulation according to any one of items 20-47.

65. Use of the amorphous solid dispersion according to any one of items1-12 and 19, or the oral dosage formulation according to any one ofitems 20-47, for preventing, treating, and/or lessening the severity ofcancer in a subject.

66. Use of the amorphous solid dispersion according to any one of items1-12 and 19, or the oral dosage formulation according to any one ofitems 20-47, for the manufacture of a medicament for preventing,treating, and/or lessening the severity of cancer in a subject.

67. The use of item 65 or 66, wherein said cancer is breast cancer,ovarian cancer, prostate cancer, cervical cancer, lung cancer, renalcancer, bladder cancer, glioma, skin cancer, head and neck carcinoma,non-Hodgkin's lymphoma, neuroblastoma, Ewing's sarcoma or Kaposi'ssarcoma.

68. Use of amorphous solid dispersion according to any one of items 1-12and 19, or the oral dosage formulation according to any one of items20-47, for preventing, treating, and/or lessening the severity of adisease or condition associated with a lipid imbalance in a subject.

69. Use of amorphous solid dispersion according to any one of items 1-12and 19, or the oral dosage formulation according to any one of items20-47, for the manufacture of a medicament for preventing, treating,and/or lessening the severity of a disease or condition associated witha lipid imbalance in a subject.

70. The use of item 68 or 69, wherein said disease or conditionassociated with lipid imbalance is Cystic Fibrosis.

71. The use of item 68 or 69, wherein said disease or conditionassociated with lipid imbalance is a respiratory disease, a neuralinjury, or a neurodegenerative disease.

72. The use of item 71, wherein said neural injury is Spinal Cord Injury(SCI)

73. The use of item 71, wherein said neurodegenerative disease isAmyotrophic Lateral Sclerosis (ALS).

74. Use of the amorphous solid dispersion according to any one of items1-12 and 19, or the oral dosage formulation according to any one ofitems 20-47, for preventing and/or treating Cystic Fibrosis or acondition associated with Cystic Fibrosis in a subject.

75. Use of the amorphous solid dispersion according to any one of items1-12 and 19, or the oral dosage formulation according to any one ofitems 20-47, for the manufacture of a medicament for preventing,treating, and/or lessening the severity of Cystic Fibrosis or acondition associated with Cystic Fibrosis in a subject.

76. Use of the amorphous solid dispersion according to any one of items1-12 and 19, or the oral dosage formulation according to any one ofitems 20-47, for preventing, treating, and/or lessening the severity ofa bone disease or condition associated with a bone disease in a subject.

77. Use of the amorphous solid dispersion according to any one of items1-12 and 19, or the oral dosage formulation according to any one ofitems 20-47, for the manufacture of a medicament for preventing,treating, and/or lessening the severity of a bone disease or conditionassociated with a bone disease in a subject.

78. The use of item 76 or 77, wherein said subject suffers from CysticFibrosis.

79. Use of the amorphous solid dispersion according to any one of items1-12 and 19, or the oral dosage formulation according to any one ofitems 20-47, for preventing, treating and/or lessening the severity of adisease or condition associated with inflammation in a subject.

80. Use of the amorphous solid dispersion according to any one of items1-12 and 19, or the oral dosage formulation according to any one ofitems 20-47, for the manufacture of a medicament for preventing,treating and/or lessening the severity of a disease or conditionassociated with inflammation in a subject.

81. The use of item 79 or 80, wherein said inflammation is lunginflammation, and the disease or condition associated with lunginflammation is Cystic Fibrosis.

82. The use of item 78 or 80, wherein said disease or conditionassociated with inflammation is a respiratory disease, a neural injury,or a neurodegenerative disease.

83. The use of item 82, wherein said respiratory disease is allergicasthma.

84. The use of item 82, wherein said neural injury is Spinal Cord Injury(SCI).

85. The use of item 82, wherein said neurodegenerative disease isAmyotrophic Lateral Sclerosis (ALS).

86. Use of the amorphous solid dispersion according to any one of items1-12 and 19, or the oral dosage formulation according to any one ofitems 20-47, for preventing, treating, and/or lessening the severity ofa disease or condition associated with opportunistic infection in asubject.

87. Use of the amorphous solid dispersion according to any one of items1-12 and 19, or the oral dosage formulation according to any one ofitems 20-47, for the manufacture of a medicament for preventing,treating, and/or lessening the severity of a disease or conditionassociated with opportunistic infection in a subject.

88. The use of item 86 or 87, wherein said disease or conditionassociated with an opportunistic infection is a respiratory disease.

89. The method of item 88, wherein said respiratory disease is CysticFibrosis.

90. Use of the amorphous solid dispersion according to any one of items1-12 and 19, or the oral dosage formulation according to any one ofitems 20-47, for preventing, treating, and/or lessening the severity ofa disease in a subject, wherein the disease is allergic asthma, SpinalCord Injury, Amyotrophic Lateral Sclerosis, diabetes, obesity, maculardegeneration, AIDS, allergic encephalomyelitis, ichthyosis, or a viralinfection caused by a flavivirus.

91. Use of the amorphous solid dispersion according to any one of items1-12 and 19, or the oral dosage formulation according to any one ofitems 20-47, for the manufacture of a medicament for preventing,treating, and/or lessening the severity of a disease in a subject,wherein the disease is allergic asthma, Spinal Cord Injury, AmyotrophicLateral Sclerosis, diabetes, obesity, macular degeneration, AIDS,allergic encephalomyelitis, ichthyosis or a viral infection caused by aflavivirus.

92. The amorphous solid dispersion according to any one of items 1-12and 19, or the oral dosage formulation according to any one of items20-47, for use in the prevention and/or treatment of cancer in asubject.

93. The amorphous solid dispersion according to any one of items 1-12and 19, or the oral dosage formulation according to any one of items20-47, for use in the manufacture of a medicament for the preventionand/or treatment of cancer in a subject.

94. The amorphous solid dispersion or oral dosage formulation for useaccording to item 92 or 93, wherein said cancer is breast cancer,ovarian cancer, prostate cancer, cervical cancer, lung cancer, renalcancer, bladder cancer, glioma, skin cancer, head and neck carcinoma,non-Hodgkin's lymphoma, neuroblastoma, Ewing's sarcoma or Kaposi'ssarcoma.

95. The amorphous solid dispersion according to any one of items 1-12and 19, or the oral dosage formulation according to any one of items20-47, for use in preventing, treating, and/or lessening the severity ofa disease or condition associated with a lipid imbalance in a subject.

96. The amorphous solid dispersion according to any one of items 1-12and 19, or the oral dosage formulation according to any one of items20-47, for use in the manufacture of a medicament for preventing,treating, and/or lessening the severity of a disease or conditionassociated with a lipid imbalance in a subject.

97. The amorphous solid dispersion or oral dosage formulation for useaccording to item 95 or 96, wherein said disease or condition associatedwith lipid imbalance is Cystic Fibrosis.

98. The amorphous solid dispersion or oral dosage formulation for useaccording to item 95 or 96, wherein said disease or condition associatedwith lipid imbalance is a respiratory disease, a neural injury, or aneurodegenerative disease.

99. The amorphous solid dispersion according to any one of items 1-12and 19, or the oral dosage formulation according to any one of items20-47, for use in the prevention, treatment, and/or lessening theseverity of Cystic Fibrosis or a condition associated with CysticFibrosis in a subject.

100. The amorphous solid dispersion according to any one of items 1-12and 19, or the oral dosage formulation according to any one of items20-47, for use in the manufacture of a medicament for preventing,treating, and/or lessening the severity of Cystic Fibrosis or acondition associated with Cystic Fibrosis in a subject.

101. The amorphous solid dispersion according to any one of items 1-12and 19, or the oral dosage formulation according to any one of items20-47, for use in preventing, treating, and/or lessening the severity ofa bone disease or condition associated with a bone disease in a subject.

102. The amorphous solid dispersion according to any one of items 1-12and 19, or the oral dosage formulation according to any one of items20-47, for use in the manufacture of a medicament for preventing,treating, and/or lessening the severity of a bone disease or conditionassociated with a bone disease in a subject.

103. The amorphous solid dispersion or oral dosage formulation for useaccording to item 102 or 103, wherein said disease or conditionassociated with bone disease is Cystic Fibrosis.

104. The amorphous solid dispersion according to any one of items 1-12and 19, or the oral dosage formulation according to any one of items20-47, for use in preventing, treating and/or lessening the severity ofa disease or condition associated with inflammation in a subject.

105. The amorphous solid dispersion according to any one of items 1-12and 19, or the oral dosage formulation according to any one of items20-47, for use in the manufacture of a medicament for preventing,treating, and/or lessening the severity of a disease or conditionassociated with inflammation in a subject.

106. The amorphous solid dispersion or oral dosage formulation for useaccording to item 104 or 105, wherein said inflammation is lunginflammation, and the disease or condition associated with lunginflammation is Cystic Fibrosis.

107. The amorphous solid dispersion or oral dosage formulation for useaccording to item 104 or 105, wherein said disease or conditionassociated with inflammation is a respiratory disease, a neural injury,or a neurodegenerative disease.

108. The amorphous solid dispersion or oral dosage formulation for useaccording to item 107, wherein said respiratory disease is allergicasthma.

109. The amorphous solid dispersion or oral dosage formulation for useaccording to item 107, wherein said neural injury is Spinal Cord Injury(SCI).

110. The amorphous solid dispersion or oral dosage formulation for useaccording to item 107, wherein said neurodegenerative disease isAmyotrophic Lateral Sclerosis (ALS).

111. The amorphous solid dispersion according to any one of items 1-12and 19, or the oral dosage formulation according to any one of items20-47, for use in preventing, treating, and/or lessening the severity ofa disease or condition associated with opportunistic infection in asubject.

112. The amorphous solid dispersion according to any one of items 1-12and 19, or the oral dosage formulation according to any one of items20-47, for use in the manufacture of a medicament for preventing,treating, and/or lessening the severity of a disease or conditionassociated with opportunistic infection in a subject.

113. The amorphous solid dispersion or oral dosage formulation for useaccording to item 111 or 112, wherein said disease or condition isassociated with an opportunistic infection is a respiratory disease.

114. The amorphous solid dispersion or oral dosage formulation for useaccording to item 113, wherein said respiratory disease is CysticFibrosis.

115. The amorphous solid dispersion according to any one of items 1-12and 19, or the oral dosage formulation according to any one of items20-47, for use in preventing, treating, and/or lessening the severity ofa disease in a subject, wherein the disease is allergic asthma, SpinalCord Injury, Amyotrophic Lateral Sclerosis, diabetes, obesity, maculardegeneration, AIDS, allergic encephalomyelitis, ichthyosis or a viralinfection caused by a flavivirus.

116. The amorphous solid dispersion according to any one of items 1-12and 19, or the oral dosage formulation according to any one of items20-47, for use in the manufacture of a medicament for preventing,treating, and/or lessening the severity of a disease in a subject,wherein the disease is allergic asthma, Spinal Cord Injury, AmyotrophicLateral Sclerosis, diabetes, obesity, dry-form age-related maculardegeneration, Stargardt Disease, AIDS, allergic encephalomyelitis,ichthyosis or a viral infection caused by a flavivirus.

Other objects, advantages and features of the present invention willbecome more apparent upon reading of the following non-restrictivedescription of specific embodiments thereof, given by way of exampleonly with reference to the accompanying drawings.

BRIEF DESCRIPTION OF DRAWINGS

In the appended drawings:

FIG. 1 shows XRPD Diffractograms of Fenretinide spray-dried intermediate(SDI) lots L215-01005a, b and c. Fenretinide lot C00324 (Fenretinide asreceived) was used as a reference.

FIG. 2 shows the pharmacokinetics profiles of 6 prototype formulationsin rats after oral dose of 10 mg.

FIG. 3 shows 4-HPR (fenretinide) plasma profiles of 6 formulations aftersingle oral dose in rats.

FIG. 4 shows 4-MPR (N-[4-methoxyphenyl]retinamide) plasma profiles of 6formulations after single oral dose of 4-HPR in rats.

FIG. 5 shows plasma exposure (AUC₀₋₄₈) of 4-HPR and 4-MPR of 6formulations (groups 7 to 12) after single oral dose in rats. Leftbars=fenretinide (4-HPR); right bars=MPR.

FIG. 6 shows SEM micrographs of A) Fenretinide lot C00324 (as received,reference), Fenretinide/PKV K30 40/60% w/w lots: B) L215-01009A, C)L215-01009B, D) L215-01009C, E) L215-01010 and F) L215-01011 at 500×.

FIG. 7 shows XRPD diffractograms of Fenretinide/PVP K30 40/60% w/w SDIlots L215-01009 to L215-01011. Fenretinide lot C00324 (as received) asreference.

FIG. 8 shows a DSC thermogram of Fenretinide/PVP K30 40/60% w/w SDI lotsL215-01009 to L215-01011. Fenretinide lot C00324 (as received) asreference.

FIG. 9 shows a TGA thermogram of Fenretinide/PVP K30 40/60% w/w SDI lotsL215-01009 to L215-01011. Fenretinide lot C00324 (as received) asreference.

FIG. 10 shows the dissolution profile of L215-01012B capsule formulationincubated at 5° C., 25° C./60% RH and 40° C./75% RH closed cap for up to1 month.

FIG. 11 shows XRPD diffractograms of Fenretinide/PVP K30 40/60% w/w SDIlot L215-01011 incubated at 5° C., 25° C./60% RH and 40° C./75% RHclosed cap for 1 month. Fenretinide lot C00324 (as received) asreference.

FIG. 12 shows XRPD diffractograms of Fenretinide 100 mg HGC lotL215-01012B incubated at 5° C., 25° C./60% RH and 40° C./75% RH closedcap for 1 month. Fenretinide lot C00324 (as received) and PlaceboCapsule lot L215-01013P as reference.

FIG. 13 shows XRPD diffractograms of Fenretinide SDIs aftermanufacturing (T=0) and after 3.5 months of storage at 5° C. (bulkpowder in amber glass bottles).

FIG. 14 shows XRPD diffractograms of Fenretinide/PVP K30 40/60% w/w SDIlot L215-01011 incubated at 5° C., 25° C./60% RH and 40° C./75% RH.Fenretinide lot C00324 (As Received) as Reference.

FIG. 15 shows XRPD diffractograms of Fenretinide SDI lots L215-01016 toL215-01020.

FIG. 16 shows XRPD diffractograms of Fenretinide SDI lots L215-01023 toL215-01027.

FIG. 17 shows antioxidants/PVP based Fenretinide SDI Formulations (20 gof solids/lot)

FIG. 18 shows 100 mg Fenretinide Capsule (L215-01028) and Tablets(L215-01029 and 030)

FIG. 19 shows 100 mg Fenretinide Capsule (L215-01031) and Tablets(L215-01032 and 033)

FIG. 20 shows Summary Results for Assay and Related Substances for SDIlots L215-01016 to 20, T=0, 1 and 3 months.

FIG. 21 shows Description, KF, Assay and Related Substances and Yieldfor SDI lots L215-01023 to 27 at T=0

FIG. 22 shows Summary Results for Assay and Related Substances for DrugProducts lots L215-01023 to 27 at T=0, after 1 month, 3 months and 6months

DISCLOSURE OF INVENTION

In the studies described herein, the present inventors have developed aspray-dried amorphous solid dispersion of fenretinide that exhibitsimproved bioavailability, and more specifically higher fenretinideplasma AUC₍₀₋₄₈₎ and C_(max) values in a rat model, relative to thecurrent fenretinide corn-oil liquid suspension.

Accordingly, in an aspect, the present invention provides an amorphoussolid dispersion comprising fenretinide or an analog thereof and atleast one matrix polymer.

“Solid dispersion” as used herein refers to a solid material, in which adrug (fenretinide or an analog thereof) is dispersed in the solid matrixpolymer. Such solid dispersions are also referred to in the art as“molecular dispersions” or “solid solutions” of the drug in the polymer.

Solid dispersions may be obtained by various techniques, for examplefast evaporation, spray-drying, precipitation or melt extrusion (e.g.,hot melt extrusion, HME). In an embodiment, the solid dispersion isobtained by spray-drying (spray-dried solid dispersion).

“Spray-dried solid dispersion” or “spray-dried dispersion” (SDD), asused herein means a solid dispersion produced using spray-dryingtechnology. The term “spray-drying” is used conventionally and refers toprocesses involving breaking up liquid mixtures into small droplets(atomization) and rapidly removing solvent from the mixture in acontainer (spray-drying apparatus), in which there is a strong drivingforce for evaporation of solvent from the droplets. Spray-dryingprocesses and spray-drying equipment or apparatus are describedgenerally in for example Perry's Chemical Engineers’ Handbook (EighthEdition 2007). More details on spray-drying processes and equipment arereviewed by Marshall, “Atomization and Spray-Drying,” 50 Chem. Eng.Prog. Monogr. Series 2 (1954), and Masters, Spray Drying Handbook (FifthEdition 1991). The strong driving force for solvent evaporation isgenerally provided by maintaining the partial pressure of solvent in thespray-drying apparatus well below the vapor pressure of the solvent atthe temperature of the drying droplets. This is accomplished by (1)maintaining the pressure in the spray-drying apparatus at a partialvacuum (e.g., 0.01 to 0.50 atm); or (2) mixing the liquid droplets witha warm drying gas; or (3) both (1) and (2). In addition, at least aportion of the heat required for evaporation of solvent may be providedby heating the spray solution. Spray-drying processes and apparatussuitable for use in the present invention include those disclosed inU.S. Pat. Nos. 7,780,988 and 7,887,840.

In an embodiment, at least a major portion of the fenretinide or analogthereof in the dispersion is amorphous. As used herein, the term “amajor portion” of the fenretinide or analog thereof means that more than50% of the fenretinide or analog thereof (by weight) in the dispersionis in the amorphous form, as opposed to the crystalline form. Inembodiments, at least 55, 60, 65, 70, 75, 80, 85, 90% or 95% of thefenretinide or analog thereof (by weight) in the dispersion is in theamorphous form. By “amorphous” is meant that the fenretinide or analogthereof is in a non-crystalline state. Preferably, the fenretinide oranalog thereof in the dispersion is “substantially amorphous”, meaningthat the amount of the fenretinide or analog thereof in crystalline formdoes not exceed about 25%, in further embodiments does not exceed about20%, 15% or 10%. More preferably, the fenretinide or analog thereof inthe dispersion is “almost completely amorphous,” meaning that the amountof fenretinide or analog thereof in the crystalline form does not exceedabout 10%. In a further embodiment, no recognizable characteristiccrystalline fenretinide or fenretinide analog peaks are present in anX-ray powder diffraction pattern of the material. Amounts of crystallinedrug may be measured by Powder X-Ray Diffraction (PXRD) analysis,Scanning Electron Microscope (SEM) analysis, Differential Scanningcalorimetry (DSC) or any other standard quantitative measurement knownin the art.

Examples of “matrix polymers”, also referred to in the field as“concentration-enhancing polymers” or “dispersion polymers”, which maybe suitable for use in the present invention, are discussed in detail infor example U.S. Pat. Nos. 7,780,988 and 7,887,840. The matrix polymercan be any pharmaceutically acceptable polymer that once coprocessedwith fenretinide or an analog thereof, functions to maintain thefenretinide/fenretinide analog in amorphous form.

Examples of polymers that may be suitable for use with the presentinvention comprise non-ionizable (neutral) non-cellulosic polymers.Exemplary polymers include: vinyl polymers and copolymers having atleast one substituent selected from hydroxyl, alkylacyloxy, andcyclicamido; polyvinyl alcohols that have at least a portion of theirrepeat units in the unhydrolyzed (vinyl acetate) form; polyvinyl alcoholpolyvinyl acetate copolymers; polyvinyl pyrrolidone; and polyethylenepolyvinyl alcohol copolymers; and polyoxyethylene-polyoxypropylenecopolymers.

Other examples of polymers that may be suitable for use with the presentinvention comprise ionizable non-cellulosic polymers. Exemplary polymersinclude: carboxylic acid-functionalized vinyl polymers, such as thecarboxylic acid functionalized polymethacrylates and carboxylic acidfunctionalized polyacrylates such as the EUDRAGIT® series,amine-functionalized polyacrylates and polymethacrylates; proteins suchas gelatin and albumin; and carboxylic acid functionalized starches suchas starch glycolate.

Other examples polymers that may be suitable for use with the presentinvention comprise nonionizable cellulosic polymers that may be used asthe polymer include: hydroxypropyl methyl cellulose acetate,hydroxypropyl methyl cellulose (HPMC), hydroxypropyl cellulose, methylcellulose, hydroxyethyl methyl cellulose, hydroxyethyl celluloseacetate, hydroxyethyl ethyl cellulose, and the like.

While specific polymers have been discussed as being suitable for use inthe dispersions formable by the present invention, blends of suchpolymers may also be suitable. Thus, the term “matrix polymer” isintended to include blends of polymers in addition to a single speciesof polymer.

In an embodiment, the matrix polymer comprises polyvinylpyrrolidone. Inanother embodiment, the matrix polymer is a polyvinylpyrrolidone, forexample polymers sold under the trade-name Plasdone® (povidones),polyvinylpyrrolidone K12, polyvinylpyrrolidone K17, polyvinylpyrrolidoneK25, polyvinylpyrrolidone K30 or polyvinylpyrrolidone K90.

In an embodiment, the ratio of fenretinide or analog thereof/matrixpolymer is from about 1:5 to about 5:1, in further embodiments about 1:4to about 4:1, about 1:3 to about 3:1, about 1:2 to about 2:1 or about1.5:1 to about 1:1.5, by weight. In an embodiment, the solid dispersioncomprises between about 30 to about 50% of fenretinide or analogthereof, and between about 50 to about 70% of matrix polymer. In anotherembodiment, the solid dispersion comprises between about 40% offenretinide or analog thereof, and about 60% of matrix polymer, byweight.

In an embodiment, the solid dispersion comprises one or more additives.Additives that may be suitable for use with the present inventioncomprise antioxidant agents. Exemplary antioxidants include: L-ascorbicacid (vitamin C), propyl gallate, sodium sulfite, sodium metabisulfite,sodium bisulfite, thioglycerol, thioglycollic acid, tocopherols andtocotrienols, butylated hydroxyanisole (BHA), butylated hydroxytoluene(BHT) or any combination thereof. In an embodiment, the matrix polymeror solid dispersion comprises BHA and/or BHT as antioxidant agent(s). Inan embodiment, the matrix polymer or solid dispersion comprises BHA andBHT as antioxidant agents. In an embodiment, the matrix polymercomprises L-ascorbic acid as antioxidant agent. In an embodiment, theantioxidant agent(s) is/are present in an amount of about 0.01% to about5%, in further embodiments in an amount of about 0.1% to about 5%, about0.2% to about 4%, 0.5% to about 3% or 0.5% to about 2%.

Fenretinide (all-trans-N-(4-hydroxyphenyl) retinamide; also referred toas 4-HPR, retinoic acid p-hydroxyanilide), which has CAS registry number65646-68-6, is a synthetic retinoid of the following formula:

Functional analogs (and/or metabolites) of fenretinide (i.e. whichexhibit the same biological activity as fenretinide) may also be used inthe present invention. As used herein, a “fenretinide analog” or“fenretinide analog/metabolite” refers to a compound that shares certainchemical structural features with fenretinide but at the same timecomprises one or more modifications thereto, and which exhibits similarbiological activity as fenretinide (but may exhibit such activity to adifferent extent). Examples of analogs or analogs/metabolites offenretinide that may be used include, but are not limited to,4-oxo-N-(4-hydroxyphenyl)retinamide (4-oxo-4-HPR),N-(4-methoxyphenyl)retinamide (4-MPR), 4-Hydroxybenzylretinone,C-glycoside and arylamide analogues of N-(4-hydroxyphenyl)retinamide-O-glucuronide, including but not limited to4-(retinamido)phenyl-C-glucuronide, 4-(retinamido)phenyl-C-glucoside,4-(retinamido)benzyl-C-xyloside; and retinoyl β-glucuronide analoguessuch as, for example, 1-(β-D-glucopyranosyl) retinamide,1-(D-glucopyranosyluronosyl) retinamide and bexarotene, described in WO07/136636, U.S. Patent Application No. 2006/0264514, U.S. Pat. Nos.5,516,792, 5,663,377, 5,599,953, 5,574,177, Anding et al. (2007) CancerRes. 67: 6270-6277 and Bhatnagar et al. (1991) Biochem. Pharmacol. 41:1471-7. In an embodiment, the fenretinide/fenretinide analog isrepresented by formula I

R is OH, COOH, CH₂OH, CH₂CH₂OH, or CH₂COOH;

carbons a-d and f-i are optionally substituted with one or more groupsselected from CH₃, OH, COOH, (CH₃)₂ and CH₂OH, or any combinationthereof, and

carbon e is optionally substituted with a C₁-C₃ alkyl group that isoptionally substituted with CH₃ and/or OH.

In an embodiment, the solid dispersion comprises fenretinide or apharmaceutically acceptable salt thereof. In a further embodiment, thesolid dispersion comprises fenretinide.

Preparation of Solid Dispersions

Dispersions of the fenretinide or an analog thereof and matrix polymermay be made via any process/technique that results in the fenretinide oranalog thereof (preferably at least a major portion, i.e., more than50%) being in the amorphous state of the fenretinide or analog thereofbeing in the amorphous state. Examples of such processes include fastevaporation, spray-drying, precipitation or melt extrusion (e.g., hotmelt extrusion, HME). In an embodiment, the solid dispersion is made byspray-drying.

Spray-drying processes and spray-drying equipment are describedgenerally in for example Perry's Chemical Engineers' Handbook (EighthEdition 2007), Marshall, “Atomization and Spray-Drying,” 50 Chem. Eng.Prog. Monogr. Series 2 (1954), and Masters, Spray Drying Handbook (FifthEdition 1991).

The dispersions generally have their maximum bioavailability andstability when the drug (fenretinide or analog thereof) is dispersed inthe matrix polymer such that it is substantially amorphous andsubstantially homogeneously distributed throughout the polymer. Ingeneral, as the degree of homogeneity of the dispersion increases, theenhancement in the aqueous concentration of the fenretinide or analogthereof and relative bioavailability increases as well. Thus, mostpreferred are dispersions having a single glass transition temperature,which indicates a high degree of homogeneity.

In the spray-drying process, the fenretinide or analog thereof and oneor more matrix polymers are dissolved in a common solvent. “Common” heremeans that the solvent, which can be a mixture of compounds, willdissolve the fenretinide or analog thereof and the polymer(s). Anantioxidant or a combination thereof, such as L-ascorbic acid, BHAand/or BHT, may be added to the mixture, for example to stabilize thechemical integrity of fenretinide against degradation by oxidation.After both the fenretinide or analog thereof and the polymer have beendissolved, the solvent is rapidly removed by evaporation in thespray-drying apparatus, resulting in the formation of a substantiallyhomogeneous, amorphous solid dispersion. In such substantiallyhomogeneous dispersions, the fenretinide or analog thereof is dispersedas homogeneously as possible throughout the polymer and can be thoughtof as a solid solution of fenretinide or analog thereof dispersed in thepolymer.

The solvent is removed by the spray-drying process. The termspray-drying is used conventionally and broadly refers to processesinvolving breaking up liquid mixtures into small droplets (atomization)and rapidly removing solvent from the mixture in a spray-dryingapparatus where there is a strong driving force for evaporation ofsolvent from the droplets. Such a strong driving force for solventevaporation is generally provided by maintaining the partial pressure ofsolvent in the spray-drying apparatus well below the vapor pressure ofthe solvent at the temperature of the drying droplets. This isaccomplished for example by either (1) maintaining the pressure in thespray-drying apparatus at a partial vacuum (e.g., 0.01 to 0.50 atm); (2)mixing the liquid droplets with a warm drying gas; or (3) both (1) and(2). In addition, at least a portion of the heat required forevaporation of solvent may be provided by heating the spray solution.

Solvents suitable for spray-drying can be any organic compound in whichthe fenretinide or analog thereof and matrix polymer are mutuallysoluble. Preferably, the solvent is also volatile with a boiling pointof 150° C. or less. In addition, the solvent should preferably haverelatively low toxicity and be removed from the dispersion to a levelthat is acceptable according to The International Committee onHarmonization (ICH) guidelines. Removal of solvent to this level mayrequire a processing step such as tray-drying or secondary dryingsubsequent to the spray-drying process. Examples of solvents includealcohols such as methanol, ethanol, n-propanol, isopropanol, andbutanol; ketones such as acetone, methyl ethyl ketone and methylisobutyl ketone; esters such as ethyl acetate and propylacetate; andvarious other solvents such as dichloromethane, acetonitrile, methylenechloride, toluene, and 1,1,1-trichloroethane. Lower volatility solventssuch as dimethylacetamide or dimethylsulfoxide can also be used.Mixtures of solvents can also be used, as can mixtures with water aslong as the polymer and fenretinide or analog thereof are sufficientlysoluble to make the spray-drying process practicable. In an embodiment,the solvent comprises dichloromethane, in a further embodiment thesolvent is 100% dichloromethane.

The composition of the solvent-bearing feed will depend on the desiredratio of drug-to-polymer in the dispersion and the solubility of thefenretinide or analog thereof and polymer in the solvent. Generally, itis desirable to use as high a combined drug and polymer concentration inthe solvent-bearing feed as possible, provided the drug and polymer aredissolved in the solvent, to reduce the total amount of solvent thatmust be removed to form the amorphous solid dispersion. Thus, thesolvent-bearing feed will generally have a combined drug and polymerconcentration of at least about 0.1 wt %, preferably at least about 1 wt%, and more preferably at least about 10 wt %. However, solvent-bearingfeeds with lower combined drug and polymer concentrations can be used toform suitable amorphous solid dispersions.

The solvent-bearing feed comprising the drug and polymer is atomizedthrough a pressure nozzle. By “pressure nozzle” is meant an atomizingmeans that produces droplets with an average droplet diameter of 50 μmor larger, with less than about 10 vol % of the droplets having a sizeless than about 10 μm. Generally, an appropriately sized and designedpressure nozzle is one that will produce droplets within this size rangewhen the spray solution is pumped through the nozzle at the desiredrate.

In the studies described herein, the solvent was evaporated using aModel GA32 Yamato® Lab Spray Dryer with the following operatingparameters: 1.2 mm nozzle; about 9-15 g/min feed rate; 1.5 kg/cm²atomization air, and 0.35-0.55 m³/min air flow. The inlet temperaturewas adjusted according to the solvent system to maintain an outlettemperature between 50-65° C. Hence, an inlet temperature of 100, 85 and70° C. was used for the lots L215-01009A, 009B and 009C, respectively.

In many cases, the solvent-bearing feed is delivered to the atomizingmeans under pressure. The pressure required is determined by the designof the atomizer, the size of the nozzle orifice, the viscosity and othercharacteristics of the solvent-bearing feed, and the desired dropletsize and size distribution. Generally, feed pressures may range from 1to 200 atm (about 0.1 to about 20 MPa) or more.

The temperature and feed rate of the drying gas is chosen so thatsufficient heat for drying the solvent-bearing feed is delivered to thedrying chamber, while allowing sufficient residence time for thedroplets to solidify before they impinge on the walls of thespray-drying apparatus. Generally, the higher the feed rate of thesolvent-bearing feed, the higher the temperature and/or flow rate of thedrying gas. Typically, the temperature of the drying gas at the inlet tothe spray dryer will be at least about 60° C. and less than about 300°C., for example between about 60 to about 100° C. In an embodiment, theinlet temperature may be adjusted according to the solvent system tomaintain an outlet temperature between about 30 to about 80° C., forexample about 50-65° C. In an embodiment, the feed rate is typically atleast about 0.1 ml/min, for example from about 1 ml/min to about 30ml/min or from about 5 ml/min to about 20 ml/min.

Following solidification, the solid powder typically stays in thespray-drying chamber for about 5 to about 60 seconds, furtherevaporating solvent from the solid powder. The final solvent content ofthe solid dispersion as it exits the dryer should be low. Generally, thesolvent content of the dispersion as it leaves the spray-drying chambershould be less than about 10 wt % and preferably less than about 3 wt %and most preferably less than about 1 wt %. As indicated above, asubsequent drying step, such as tray-drying, may be used to remove thesolvent to this level.

In an embodiment, the spray-drying process is performed under thefollowing operating parameters: 1.2 mm nozzle; about 9-15 g/min feedrate; about 1.5 kg/cm² atomization air, and about 0.35-0.55 m³/min airflow, and the inlet temperature is adjusted to maintain an outlettemperature between about 50-65° C.

In another embodiment, the spray-drying process is performed under thefollowing operating parameters: 1.2 mm nozzle; about 11-15 g/min feedrate; about 0.1-0.3 MPa atomization air, about 0.40 m³/min air flow,inlet temperature of about 75° C.; outlet temperature of about 45 toabout 48° C.

In another embodiment, the spray-drying process is performed under thefollowing operating parameters: 1.2 mm nozzle; about 20 g/min feed rate;about 0.15 MPa atomization air, about 0.45-0.48 m³/min air flow, inlettemperature of about 80° C.; outlet temperature of about 50 to about 54°C.

In certain embodiments, the material is processed though a secondarydrying step. In some embodiments, a tray dryer is used for secondarydrying. For example, in some embodiments, the dryer is a conventiondryer. The secondary drying is performed for a sufficient period of timeto meet product specifications. For example, in some embodiments,secondary drying occurs at about 30° C., 35° C., 40° C., 45° C., or 50°C. In certain embodiments, the drying time is at least about 1, 2, 3, 5,6, 7, 8, 9, or 10 hours. In certain embodiments, the drying time isabout 2 hours. In another embodiment, the secondary drying is performedunder vacuum, for example at a pressure of about −40 to about −60 kPa,e.g., about −50 kPa.

Dosage Formulations

A “dosage form” or “dosage formulation” as used herein means a unit ofadministration of an active agent. Examples of dosage formulationsinclude tablets, granules, capsules, injections, suspensions, liquids,emulsions, creams, ointments, suppositories, inhalable formulations,transdermal formulations, and the like. By “oral dosage formulation” ismeant to include a unit dosage formulation for oral administration.

In some embodiments, the amorphous solid dispersion of the presentinvention is combined with one or more optional excipients to formulatethe dispersion into suitable dosage formulations, such as tablets,capsules (e.g., hard gelatine capsules), strips, caplets, suspensions,powders for suspensions, cream, transdermal patches, depots, and thelike.

The dispersion can also be added to other dosage form ingredients in amanner that advantageously does not substantially alter the activity ofthe fenretinide or analog thereof.

Generally, excipients such as surfactants, pH modifiers, fillers, matrixmaterials, complexing agents, solubilizers, lubricants, glidants,antioxidants, and so forth may be used for customary purposes and intypical amounts without adversely affecting the properties of thecompositions. See for example, Remington's Pharmaceutical Sciences (18thed. 1990).

The addition of pH modifiers such as acids, bases, or buffers may bebeneficial, retarding the dissolution of the composition (e.g., acidssuch as citric acid or succinic acid when the matrix polymer is anionic)or, alternatively, enhancing the rate of dissolution of the composition(e.g., bases such as sodium acetate or amines when the matrix polymer iscationic).

Conventional matrix materials, complexing agents, solubilizers, fillers,diluents, disintegrating agents (disintegrants), preservatives,suspending agents or thickeners, anti-caking agents, or binders may alsobe added as part of the composition itself or added by granulation viawet or mechanical or other means. These materials may comprise up to 80or 90 wt % of the composition.

Examples of matrix materials, fillers, or diluents include, withoutlimitation, lactose, mannitol, xylitol, microcrystalline cellulose,dibasic calcium phosphate (anhydrous and dihydrate), starch, and anycombination thereof.

Examples of disintegrants include, without limitation, sodium starchglycolate, sodium alginate, carboxy methyl cellulose sodium, methylcellulose, and croscarmellose sodium, and crosslinked forms of polyvinylpyrrolidone such as those sold under the trade name CROSPOVIDONE®(available from BASF Corporation), and any combination thereof.

Examples of binders include, without limitation, methyl cellulose,microcrystalline cellulose, starch, and gums such as guar gum,tragacanth, and any combination thereof.

Examples of lubricants include, without limitation, magnesium stearate,calcium stearate, stearic acid, and any combination thereof.

Examples of glidants include, without limitation, metal silicates,silicon dioxides, higher fatty acid metal salts, metal oxides, alkalineearth metal salts, and metal hydroxides. Examples of preservativesinclude, without limitation, sulfites (an antioxidant), benzalkoniumchloride, methyl paraben, propyl paraben, benzyl alcohol, sodiumbenzoate, and any combination thereof.

Examples of suspending agents or thickeners, without limitation, includexanthan gum, starch, guar gum, sodium alginate, carboxymethyl cellulose,sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methylcellulose, polyacrylic acid, silica gel, aluminum silicate, magnesiumsilicate, titanium dioxide, and any combination thereof.

Examples of anti-caking agents or fillers, without limitation, includesilicon oxide, lactose, and any combination thereof.

Examples of solubilizers include, without limitation, ethanol, propyleneglycol, polyethylene glycol, and any combination thereof.

Examples of antioxidants include, without limitation, phenolic-basedantioxidants such as butylated hydroxyanisole (BHA), butylatedhydroxytoluene (BHT), tert-butyl-hydroquinone (TBHQ),4-hydroxymethyl-2,6-di-tert-butylphenol (HMBP),2,4,5-trihydroxy-butyrophenone (THBP), propyl gallate (PG), triamylgallate, gallic acid (GA), α-Tocopherol (vitamin E), tocopherol acetate,reducing agents such as L-ascorbic acid (vitamin C), L-ascorbylpalmitate, L-ascorbyl stearate, thioglycolic acid (TGA), ascorbylpalmitate (ASP), sulphite-based antioxidants such as sodium sulphite,sodium metabisulphite, sodium bisulphite and thioglycerol and otheragents such as disodium ethylenediamine tetraacetate (EDTA), sodiumpyrophosphate, sodium metaphosphate, methionine, erythorbic acid andlecithin, and any combination thereof. In an embodiment, the formulationcomprises a combination of antioxidants. In an embodiment, theformulation comprises a combination of BHA and BHT. In an embodiment,the formulation comprises ascorbic acid.

One other class of excipients is surfactants, optionally present fromabout 0 to about 10 wt %. Suitable surfactants include, withoutlimitation, fatty acid and alkyl sulfonates; commercial surfactants suchas benzalkonium chloride (HYAMINE® 1622, available from Lonza, Inc.,Fairlawn, N.J.); dioctyl sodium sulfosuccinate (DOCUSATE SODIUM,available from Mallinckrodt Spec. Chem., St. Louis, Mo.);polyoxyethylene sorbitan fatty acid esters (TWEEN®, available from ICIAmericas Inc., Wilmington, Del.; LIPOSORB® 0-20, available from LipochemInc., Patterson N.J.; CAPMUL.™. POE-0, available from Abitec Corp.,Janesville, Wis.); and natural surfactants such as sodium taurocholicacid, 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine, lecithin, andother phospholipids and mono- and diglycerides, and any combinationthereof. Such materials can be employed to increase the rate ofdissolution by, for example, facilitating wetting, or otherwise increasethe rate of drug release from the dosage form.

Other conventional excipients, including pigments, lubricants,flavorants, humectants, solution retarding agents, absorptionaccelerators, wetting agents, absorbents, and other ones well-known inthe art, may be employed in the compositions of this invention. Forexample, excipients such as pigments, lubricants, flavorants, and soforth may be used for customary purposes and in typical amounts withoutadversely affecting the properties of the compositions.

Other components commonly added to pharmaceutical compositions include,e.g., inorganic salts such as sodium chloride, potassium chloride,calcium chloride, sodium phosphate, potassium phosphate, sodiumbicarbonate; and organic salts such as sodium citrate, potassiumcitrate, sodium acetate, etc.

In an embodiment, the amorphous solid dispersion of the presentinvention is combined with a disintegrant, for example a cross-linkedsodium carboxymethylcellulose e.g., croscarmellose (Solutab®). Otherexamples of disintegrants include corn starch, potato starch, sodiumcarboxymethylcellulose, sodium starch glycolate, sodium croscarmellose,crospovidone, and any combination thereof. In an embodiment, thedisintegrant is present in an amount from about 2 to about 10% byweight, for example from about 3 to about 8% or about 4 to about 6% byweight.

In an embodiment, the amorphous solid dispersion of the presentinvention is combined with a lubricant, for example magnesium stearate.Other examples of lubricants include talc, silicon dioxide, stearicacid, and sodium stearyl fumarate. In an embodiment, the lubricant ispresent in an amount from about 0.5 to about 2% by weight, for examplefrom about 0.8 to about 1.2% or about 1% by weight.

In an embodiment, the amorphous solid dispersion of the presentinvention is combined with a filler or diluent, for examplemicrocrystalline cellulose (Avicel®, such as Avicel®PH-102) and/orcalcium hydrogen phosphate dehydrate (Encompress®). Other examples offillers or diluents include crystalline cellulose, cellulosederivatives, acacia, corn starch, lactose, mannitol, sugars, calciumphosphate, calcium carbonate, gelatins, and any combination thereof. Inan embodiment, the filler or diluent is present in an amount from about20 to about 45% by weight, for example from about 30% to about 40% byweight, e.g., about 35%.

The amorphous solid dispersion of the present invention may be used in awide variety of dosage forms for administration by a wide variety ofroutes, including, but not limited to, oral, nasal, rectal, vaginal,transdermal, buccal, subcutaneous, intravenous, and pulmonary.

In certain embodiments, the amorphous solid dispersion as disclosedherein is formulated as an oral dosage formulation. Formulationssuitable for oral administration may be in the form of capsules,cachets, pills, tablets, lozenges (using a flavored basis, usuallysucrose and acacia or tragacanth), powders, granules, or as a solutionor a suspension in an aqueous or non-aqueous liquid, or as an elixir orsyrup, or as pastilles (using an inert matrix, such as gelatin andglycerin, or sucrose and acacia), and the like, each containing apredetermined amount of an active ingredient. A composition may also beadministered as a bolus, electuary, or paste. In an embodiment, the oraldosage formulation of the present invention is a tablet. A tablet may bemade by compression or molding, optionally with one or more accessoryingredients. Compressed tablets may be prepared using binder, lubricant,inert diluent, preservative, disintegrant, surface-active or dispersingagent. Molded tablets may be made by molding in a suitable machine amixture of the powdered inhibitor(s) moistened with an inert liquiddiluent.

Tablets, and other solid dosage forms, such as dragees, capsules, pills,and granules, may optionally be scored or prepared with coatings andshells, such as enteric coatings and other coatings well known in thepharmaceutical-formulating art. They may also be formulated so as toprovide slow or controlled release of the active ingredient thereinusing, for example, hydroxypropylmethyl cellulose in varying proportionsto provide the desired release profile, other polymer matrices,liposomes, and/or microspheres. They may be sterilized by, for example,filtration through a bacteria-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved in sterile water, or some other sterile injectable mediumimmediately before use. These compositions may also optionally containopacifying agents and may be of a composition that they release theactive ingredient(s) only, or preferentially, in a certain portion ofthe gastrointestinal tract, optionally, in a delayed manner.

In some embodiments of the oral dosage formulation as disclosed herein,the amorphous solid dispersion is present in an amount of from about 10to about 90%, about 20 to about 80%, about 30 to about 60% or about 45to about 55% by weight, or another range within the values providedherein.

In an embodiment, the amorphous solid dispersion or the dosageformulation of the present invention results in fenretinide (or ananalog thereof) plasma AUC₍₀₋₄₈₎ and/or C_(max) values that are at least1.5-times (in further embodiments at least 2-, 2.5-, 3-, or 4-times)higher relative to the normalized fenretinide plasma AUC₍₀₋₄₈₎ and/orC_(max) values exhibited by a corresponding fenretinide corn oil liquidsuspension, e.g., AUC₍₀₋₄₈₎ and/or C_(max) values measured in an animalmodel, such as rats.

Uses of the Amorphous Solid Dispersion and the Dosage Formulation

The amorphous solid dispersion and the dosage formulation as disclosedherein may be used for preventing or treating (e.g., alleviating) one ormore symptoms and/or severity of any disease/condition that is subjectto prevention or treatment by administering fenretinide or a fenretinideanalog. For example, conditions that may be prevented or treated by thedispersion or dosage form of the present invention includehyperproliferative disorders, malignancies and neoplasms (e.g., solidtumors, cancers), such as those disclosed in WO 2002/058689. Suchhyperproliferative disorders, malignancies, and neoplasms include, butare not limited to, malignant disorders such as breast cancers;osteosarcomas; angiosarcomas; fibrosarcomas and other sarcomas (e.g.,Ewing's sarcoma, Kaposi's sarcoma); leukemias; lymphomas (e.g.,non-Hodgkin's lymphoma); sinus tumors; ovarian, uretal, bladder,prostate and other genitourinary cancers; colon esophageal and stomachcancers and other gastrointestinal cancers; lung cancers (non-small celllung cancers); myelomas; pancreatic cancers; liver cancers; kidneycancers; endocrine cancers; skin cancers (e.g., melanoma, basal cellcarcinoma); head and neck carcinoma and brain or central and peripheralnervous (CNS) system tumors, malignant or benign, including gliomas andneuroblastomas.

Examples of diseases/conditions that are subject to prevention ortreatment by administering fenretinide include also premalignant andnon-neoplastic or non-malignant hyperproliferative disorders such asmyelodysplastic disorders; cervical carcinoma-in-situ; familialintestinal polyposes such as Gardner syndrome; oral leukoplakias;histiocytoses; keloids; hemangiomas; hyperproliferative arterialstenosis, inflammatory arthritis; hyperkeratoses and papulosquamouseruptions including arthritis. Also included are viral-inducedhyperproliferative diseases such as warts and Epstein-Barr virus(EBV)-induced disease (i.e., infectious mononucleosis), scar formation,and the like.

Other diseases/conditions may be prevented or treated by the amorphoussolid dispersion or dosage form of the present invention include thosediscussed in PCT Patent Publication Nos. WO 2005/084657, WO 2007/068116and WO 2009/103106, for example conditions associated with inflammationof the respiratory tract such as cystic fibrosis, allergic asthma, adisease or condition associated with a lipid or fatty acid imbalance(DHA/AA imbalance), including infections (e.g., opportunisticinfections) of the respiratory tract (e.g., Haemophilus influenzae,Pseudomonas aeruginosa, Streptococcus pneumoniae, Streptococcuspyogenes, Mycobacterium tuberculosis, Candida albicans or Aspergillusfumigatus) and bone diseases (osteopenia or osteoporosis), as well asneural diseases or conditions associated with neuroinflammation and/ormicroglial activation, such as neural injury (e.g., spinal cord injury)and neurodegenerative diseases (e.g., Amyotrophic Lateral Sclerosis(ALS), Parkinson's disease, and Huntington's disease). Otherdiseases/conditions may be prevented or treated by the spray-driedamorphous solid dispersion or dosage form of the present inventioninclude those discussed in PCT publication No. WO 2009/114136, forexample conditions associated with HIV/AIDS, allergic encephalomyelitis,ichthyosis, and metabolic conditions such as diabetes and obesity, aswell as ophthalmic conditions described in PCT publication No.WO2012078525, such as various macular degenerations and dystrophies,including but not limited to dry-form age-related macular degeneration(dry AMD) and Stargardt Disease.

Other diseases/conditions may be prevented or treated by the amorphoussolid dispersion or dosage form of the present invention include thosediscussed in PCT publication No. WO 2014/169355, such as viralinfections caused by flaviviruses, such as dengue virus, yellow fevervirus, West Nile virus or Japanese encephalitis virus or infectionscaused by Chikungunya virus (CHIKV).

It is also contemplated that the solid dispersion and the dosageformulation of the instant invention (or combinations thereof) may beused alone or in combination with (i.e., administered before, after, orsimultaneously with) other therapeutics and/or nutraceuticals and/ornutritional supplements, currently used to prevent and/or treat theabove-noted diseases and/or conditions (e.g., cancers, cystic fibrosis,spinal cord injury or neurodegenerative diseases/disorders, metabolicdiseases/conditions, ophthalmic conditions) or their associated effects(e.g., pain). An example for such combination could include fenretinideand docosahexaenoic acid (DHA) for the prevention and/or the treatmentof disease or condition associated with AA/DHA lipid/fatty acidimbalance, such as cystic fibrosis, inflammation, opportunisticinfection, neuroinflammatory and neurodegenerative diseases.

MODE(S) FOR CARRYING OUT THE INVENTION

The present invention is illustrated in further detail by the followingnon-limiting examples.

Example 1: Materials & Methods

Formulation Approaches

Reagents

Table 1 provides a description of the materials used in this study. Allmaterials were stored at room temperature (RT).

TABLE 1 Materials Material (Commercial Name) Lot # Supplier FenretinideC00324 Cedarburg Lactose monohydrate (FastFlo ® 316) C00301 ForemostLactose monohydrate (Granulac ® 200) L1020A4172 Maggie Lactosemonohydrate (Tabletose ®-80) C00125 Meggle Miorocrystalline cellulose(Avicel ® PH-102) C00098 FMC Microcrystalline cellulose (Tabulose ®-101)C00044 Blanver Pregelatinized maize starch (Starch 1500) IN518955Colorcon Calcium hydrogen phosphate dihydrate (Emcompress ®) C00084 JRSSodium lauryl sulfate (SLS) C00038 Stepanol WA-100 Poloxamer ® 188C00261 BASF Craspovidone CL 18682656PO BASF Microencapsulatedpolisorbate 80 (Sepitrap ® 80) 249081 SEPPIC Croscarmellose sodium(Solutab ® Type A) C00020 Blanver Colloidal silicon dioxide (Aerosil ®200) C00122 Evonik Magnesium stearate (Ligamed ® MF-2-V) C00124 PeterGraven Lauroyl polyoxyl-32 glycerides (Gelucire ® 44/14) 125008Gattefosse Macrogol 15 hydroxystearate (Kolliphor ® HS 15) 50383647G0BASF Polyethylene glycol 400 20738568 A&C Hypromellose (Vivapharm ® HPMCE5) 10056/10X JRS Povidone (Plasdone ® K-29/32) C00033 ISP Povidone(Kollidon ® 30) C00286 BASF Copovidone (Kollidon ® VA64) 17250416K0 BASFDichloromethane (DCM) 53102 EMD 53130 Methanol (MeOH) 53088 EMD Ethanol,anhydrous (EtOH) E00515 Commercial Alcohol Hard gelatin capsules # 00white opaque 70934661/70502051 Capsugel Hard gelatin capsules # 00orange opaque C00159 Capsugel Hard gelatin capsules # 1 white opaqueC00023 Capsugel Hard gelatin capsules # 9 white opaque 2757 CapsugelFormulation Approaches

Dry blending. L215-01001 and 002 (Table 2) is a dry-blend powderformulation of Fenretinide (40%). First, all ingredients were screenedwith a 30 mesh-sieve and mixed without the lubricant using a V-blenderat 25 rpm for 3 min. The lubricant was added and mixed for 2 min. Thefinal blend was encapsulated for an equivalent of 5 and 100 mgFenretinide/capsule.

TABLE 2 Dry Blending Formulation for Lots L215-01001 and 2 L215-01001L215-01002 Ingredients % w/w Fenretinide 40.0 FastFlo ® 316 33.8Avicel ® PH-102 11.2 Starch 1500 5.0 — Sepitrap ® 80 5.0 — Solutab ®Type A 4.0 — SLS — 5.0 Poloxamer ® 188 — 5.0 Crospovidone CL — 4.0Magnesium stearate  1.0 Total: 100.0 

Melt Granulation: L215-01003 (Table 3) was prepared by melt granulationmethod. Powdered ingredients were screened through 30-mesh sieve, mixedduring 2 min using mortar/pestle and dispersed into molten Gelucire® atapproximately 50° C. The mixture was homogenized using mortar andpestle. The mass was screened through a 20-mesh sieve to obtainuniform-sized granules. The lubricant was added and mixed for 2 min. Thefinal blend was encapsulated for an equivalent of 5 mg (for PK testingin rats) and 100 mg Fenretinide/capsule.

TABLE 3 Melt Granulation Formulation for Lot L215-01003 Ingredients %w/w Fenretinide 40.0 Gelucire ® 44/14 20.0 Granulac ® 200 26.2Tabulose ®-101 8.8 Croscarmellose sodium 4.0 Magnesium stearate 1.0Total: 100.0

Lipid-Based Dispersions: Lipid-based dispersions formulations (lotsL215-01004a to 004c) are presented in Table 4. Fenretinide (40%) wasdispersed in the in the molten carrier at 60° C. The mixture wasvigorously mixed and encapsulated for an equivalent of 5 mg and 100 mgFenretinide/capsule.

TABLE 4 Lipid-Based Dispersion Formulations for Lots L215-01004a to cL215-01004a L215-01004b L215-01004c Ingredients % w/w Fenretinide 40.0Gelucire ® 44/14 60 40.0 20.0 Polyethylene glycol 400 — 20.0 20.0Kolliphor ® HS 15 — — 20.0 Total: 100.0 

Solid Dispersions: Solid dispersions of Fenretinide (activepharmaceutical ingredient, API) were obtained by spray drying. Thespray-drying solution was prepared by dissolving API/Polymer (8 g/12 g)in 400 ml of methanol/dichloromethane (50/50% v/v) system solvent. Thesolvent was evaporated using a Model GA32 Yamato® Lab Spray Dryer withthe following operating parameters: 1.2 mm nozzle; about 18 ml/min feedrate; 70° C. inlet temperature; 31-40° C. outlet temperature; 1.5 kg/cm²atomization air, and 0.40 m³/min air flow. The spray dried material wassecondary dried for 2 hours at 40° C. and −50 kPa in an Isotemp® vacuumoven model 281 A. The Spray-Dried Intermediate (SDI) (lots L215-01005ato 005c, Table 5) were encapsulated for an equivalent of 5 and 100 mgFenretinide/capsule.

TABLE 5 Solid Dispersions Formulations for Lots L215-01005a to cL215-01005a L215-01005b L215-01005c Ingredients % w/w Fenretinide 40.0HPMC E5 60.0 — — Plasdone ® K-29/32 — 60.0 — Kollidon ® VA64 — — 60.0Total: 100.0 

Dry Granulation (DG) by Slugging: Before the preparation of prototype, anew batch of SDI was manufactured (lot L215-01006b, Table 6) accordingto the same process used for lot L005b. L215-01006b (Table 6): Spraydrying process was conducted as described for lot L215-01005b.

TABLE 6 SDI Formulation for Lot L215-01006b Ingredients % (w/w)qty/batch Fenretinide 40.0 48.0 g Plasdone ® K-29/32 60.0 72.0 gMethanol/Dichloromethane (50:50 v/v) NA 1.50 L Total: 100.0  120.0 g NA:Evaporated during the process

Fenretinide prototype formulations were prepared by slugging using thespray-dried intermediate (SDI) material from lot L215-01006b.

L215-01007 (Table 7): The slugs (350-450 mg/12 mm/4-6 kp) werecompressed using a Carver® (Model C) hydraulic hand press with 12 mmround standard concave tooling. The granules were obtained by screeningthrough a 20-mesh sieve. The extragranular ingredients and granulatedmaterial were mixed for 2 minutes at 25 rpm. The lubricant was added andmixed for 2 additional minutes. The powders were encapsulated for anequivalent of 2.5 and 100 mg Fenretinide/capsule.

TABLE 7 Dry Granulation Prototype Formulations tor Lots L215-01007a to cFormulation L215-01007a L215-01007b L215-01007c mg/ mg/ mg/ Ingredientname % w/w unit % w/w unit % w/w unit Intragranular ingredientsFenretinide/ 50.0 6.3 50.0 6.3 50.0 6.3 Plasdone ® SDI (L215-01006b)Croscarmellose 2.5 0.3 2.5 0.3 — — Magnesium stearate 0.5 0.1 0.5 0.10.5 0.06 Aerosil ®-200 — — — — 0.1 0.01 Extragranular ingredientsAvicel ®-102 22.0 2.8 28.0 3.5 22.0 2.8 Tablettose-80 22.0 2.8 — — 22.02.8 Emcompress ® — — 16.0 2.0 — — Croscarmellose 2.5 0.3 2.5 0.3 2.5 0.3Magnesium stearate 0.5 0.1 0.5 0.1 0.5 0.1 Total: 100.0 12.5  100.012.5  100.0 12.5

L215-01008 (Table 8): The slugs (about 500 mg/9.0×21.5 mm/2-6 kp) werecompressed using a Rotary tableting machine SVIAC PR6 with 9.00×21.50 mmcapsule shape tooling. The granules were obtained by screening through a20-mesh sieve. The extragranular ingredients and granulated materialwere mixed for 2 minutes at 25 rpm. The lubricant was added and mixedfor 2 additional minutes. The final blend was encapsulated for anequivalent of 100 mg Fenretinide/capsule. These capsules were packagedin plastic bottles and stored at 5° C. pending further use. A placebowas prepared (L215-01008P) by dry blending and stored in powder form.

TABLE 8 Fenretinide 100 mg Capsule Formulation Lot L215-01008 by DryGranulation Ingredients % w/w mg/unit Intragranular ingredientsFenretinide/Plasdone ® SDI (L215-01006b) 50.0 250.0 Croscarmellose 2.512.5 Magnesium stearate 0.5 2.5 Extragranular ingredients Avicel ®-10228.0 140.0 Emcompress ® 16.0 80.0 Croscarmellose 2.5 12.5 Magnesiumstearate 0.5 2.5 Total: 100.0 500.0Solid Dispersion Improvement/OptimizationSolvent System Optimization

The solubility of Fenretinide/PVP K30 40/60% w/w mixtures was firstvisually assessed in various ratios of ethanol/dichloromethane.Thereafter, Spray Dryed Intermediates (SDIs) were produced fromFenretinide/PVP K30 40/60% w/w mixture solubilized at 7.5% w/v inethanol/dichloromethane 90/10 v/v (L215-01009A), 50/50 v/v (L215-01009B)and 10/90 v/v (L215-01009C) systems for a total batch size of 15 g. Thesolvent was evaporated using Model GA32 Yamato® Lab Spray Dryer with thefollowing operating parameters: 1.2 mm nozzle; about 9-15 g/min feedrate; 1.5 kg/cm² atomization air, and 0.35-0.55 m³/min air flow. Theinlet temperature was adjusted according to the solvent system tomaintain an outlet temperature between 50-65° C. Hence, an inlettemperature of 100, 85 and 70° C. was used for the lots L215-01009A,009B and 009C, respectively.

Solvent System Optimization

Successive amounts of Fenretinide and PVP K30 40/60% w/w were added in200 ml of dichloromethane until saturation of the solution. At a totalsolid loading of 20% w/v (40 g), the addition of Fenretinide and PVP K30was stopped due to the increase of the solution viscosity. The solutionwas spray dried (lot L215-01010) with the following process parameters:11-15 g/min feed rate; 75° C. inlet temperature; 45-48° C. outlettemperature; 0.1-0.3 MPa atomization air, and 0.40 m³/min air flow.

Scale-Up

Lot L215-01011 was prepared by dissolving 100 g Fenretinide and 150 gPVP K30 in 2000 ml of dichloromethane. The solution was spray dried withthe following process parameters: about 20 g/min feed rate; 80° C. inlettemperature; 50-54° C. outlet temperature; 0.15 MPa atomization air, and0.45-0.48 m³/min air flow. At about each 1000 g of solution sprayed, thespray drying process was stopped and the SDI in the product vessel wascollected. The spray dried material was secondary dried for 16 hours atRT and −15 inHg in an Isotemp® vacuum oven model 281A.

SDI/Excipient Direct Encapsulation

SDI/excipient blend lots L215-01012A and B (batch size of 125 and 150capsules, Table 9) was encapsulated directly into 00 HGC using asemi-automatic Schaefer® STI-10 capsule filling machine. The SDI,Avicel®-102 and Emcompress® were mixed for 5 min at 25 rpm in a 1.0 qtV-blender. The blend was screened over a 600 μm sieve and returned intothe V-blender. The croscarmellose, previously screened over 600 μmsieve, was added into the V-blender and mixed for 5 min at 25 rpm.Finally, magnesium stearate, previously screened over 600 μm sieve, wasadded into the V-blender and mixed for 2 min at 25 rpm. The capsulebodies were filled manually. For analysis purposes, a placeboformulation lot L215-01013P (Table 10) was prepared as described abovefor the lot L215-01012.

TABLE 9 Fenretinide 100 mg HGC Direct Encapsulation Formulation (LotsL215-01012A and 012B) L215-01012A L215-01012B Ingredients % w/w mg/unit% w/w mg/unit Fenretinide/PVP K30 SDI 50.0 250.0 55.55 250.0(L215-010011) Avicel ®-102 17.65 75.0 17.78 80.0 Emcompress ® 17.65 75.020.0 90.0 Croscarmellose 4.9 20.75 5.67 25.5 Magnesium stearate 1.0 4.251.0 4.5 Total: 100.0 425.0 1.00 450.0

TABLE 10 Fenretinide Placebo HGC Direct Encapsulation Formulation (LotL215-01013P) L215-01013P Ingredients % w/w mg/unit PVP K30 42.86 192.86Avicel ®-102 22.86 102.86 Emcompress ® 25.71 115.71 Croscarmellose 7.2932.79 Magnesium stearate 1.29 5.79 Total: 100.0 450.0Short Term Stability Study

The short term stability of formulation lot L215-01012B was initiated byputting samples into sealed HDPE bottle at 5° C., 25° C./60% RH and 40°C./75% RH. At each time point (0.5 and 1 month), the samples wereevaluated in term of assay and related substance, moisture (KarlFisher), crystal state (XRPD) and dissolution profile.

API, SDI and Prototypes Characterization

The API, SDI and prototypes produced during this study werecharacterized by evaluation the applicable following properties:

-   -   Assay and related substance;    -   Crystal state (X-ray powder diffraction, XRPD);    -   Differential scanning calorimetry (DSC);    -   Dissolution;    -   Residual solvent;    -   Scanning electron microscopy (SEM);    -   Thermogravimetric analysis (TGA); and    -   Viscosity        Assay and Related Substance

Fenretinide assay and related substance was quantified by HPLC using thefollowing system:

HPLC System:

Equilibrate the HPLC column for at least 30 min before the run

-   -   Column: Inertsil® ODS-2, 250×4.6 mm, 5 μm    -   Column Temperature: 25° C.    -   Tray Temperature: 20° C.    -   Injection volume: 5 μL    -   Mobile Phase: A: TFA/H20 buffer pH 3.0: ACN 20:80; B:CAN (see        gradient)    -   Flow: 2.0 mL/min    -   Detector wavelength: 360 nm    -   Run Time: 35 minutes    -   Retention Time: ˜8 minutes (Fenretinide)    -   Sample Diluent: 80% Acetonitrile in water        Gradient:

Time (min.) Mobile Phase A (%) Mobile Phase B (%) 0 100 0 12 100 0 20 0100 25 0 100 25.1 100 0 35 100 0Crystal State (X-Ray Powder Diffraction, XRPD) (USP <941>, USPharmacopeia XXXIV, US Pharmacopeia Convention, Rockville, Md., 2011)

The crystal state was studied by X-Ray Diffraction (XRD) using a Bruker®D2 Phaser X-ray diffractometer with Lynxeye® detector, Cu Kα radiation(λ=1.5406 Å) at an increment of 0.04° 2θ with a 0.1 s step time(scanning rate of 24 2θ/min) over a range of 5-40° 2θ, a 1.0 mm openingslit and a 8 mm detector window. The samples (about 0.2 g) were analysedusing a low volume sample holder kept under a constant rotation of 15rpm during the analysis.

Differential Scanning Calorimetry (DSC) (USP <891>, US PharmacopeiaXXXIV, US Pharmacopeia Convention, Rockville, Md., 2011)

Differential Scanning calorimetry (DSC) analysis was completed with a TAInstrument Q20 DSC. The sample was first equilibrated at 20° C. for 5min and heated at 10° C./min up to 250° C. under a nitrogen purge of 50ml/min. The sample was analysed using a TA Instrument Tzero® hermeticaluminium pan.

Dissolution

Fenretinide dissolution profile form the prototype was characterisedusing USP apparatus II (paddle, 100 rpm, 37° C.) and HPLC using thesystems below. At 60 min a 200 rpm ramp was applied for 15 min. Thedissolution medium (900 mL) was 0.1N HCl with 2% SDS, pH 6.8 phosphatebuffer with 2% SLS or pH 8.0 phosphate buffer with 2% SDS.

Dissolution System:

Use a calibrated bath set to these conditions with 6 vessels:

-   -   Medium: 900 ml    -   Bath Temperature: 37.0±0.5° C.    -   Apparatus: USP apparatus II (Paddles)    -   Speed: 100 rpm    -   Sampling times: 10, 20, 30, 60 minutes+ramp at 200 rpm for 15        minutes    -   Sampling volume: 1 ml    -   Filter: 45 μm Polyethylene

HPLC System:

-   -   Column: Inertsil® ODS-2, 250×4.6 mm, 5 μm    -   Column Temperature: 25° C.    -   Tray Temperature: 20° C.    -   Injection volume: 5 μL    -   Mobile Phase: Isocratic 30% Mobile phase A/70% mobile phase B    -   Flow: 2.0 mL/min    -   Detector wavelength: 360 nm    -   Run Time: 6 minutes    -   Retention Time: ˜4 minutes (Fenretinide)        Residual Solvent

The amount of residual solvent in Fenretinide SDI or prototype wasquantified by gas chromatography using the system below (based on USP<467> Residual Solvents).

Equipment:

Column: DB-624, 30 m×0.32 mm, 1.8 μm film thickness. Mode: Constantflow. Column flow: 3.0 mL/min (linear velocity of 44 cm/s).

Oven program: Initial temperature: 35° C. Initial Hold time: 5.0minutes. Ramp Program #1: 30° C./min. up to 260° C., hold time 15 min.

Detector (FID): Temperature: 260° C. Hydrogen flow: 30 mL/min. Air flow:300 mL/min. Makeup: Nitrogen at 30 mL/min.

Inlet (Split/Splitless): Mode: Splitless; Vent flow of 224 mL/min @ 0.10min. Temperature: 260° C. Carrier Gas: Helium. Gas saver parameters:15.0 mL/min after 2.00 min.

Injector Parameters: Rinse Solvents: A=DMSO; B=Methylpyrrolidone.Pre-injection wash: 3× Solvent Band 2× Sample. Sample Injection: 3×Syringe pumps then 1 μL Injection volume. Post-injection wash: 10×Solvent A and 10× Solvent B.

Scanning Electron Microscopy (SEM)

The morphology and surface characteristics of particles were examined atvarious magnifications with a JEOL JSM-6010LV InTouchScope® scanningelectron microscope using a backscattered electron (BSE) or a secondaryelectron (SEI) detector. The images were obtained with acceleratingvoltages between 1.5 and 20.0 kV under a pressure of 60 to 70 Pa.Samples were placed on metallic stubs using double-sided carbonconductive tape.

Thermogravimetric Analysis (TGA) (USP <891>, US Pharmacopeia)(XXIV, USPharmacopeia Convention, Rockville, Md., 2011)

Thermogravimetric analysis was performed using a TA Instrument Q50thermogravimetric analyzer at scanning speed of 10° C./min over atemperature range of 20 to 500° C. The samples were heated in a platinumopen crucible in nitrogen atmosphere (60 ml/min) and the mass loss as afunction of temperature was recorded.

Viscosity

The viscosity of the solution was determined at RT using a Brookfield®DV-II+viscometer at 100 rpm.

Example 2: Encapsulation

Fenretinide 2.5 and 5.0 mg prototype formulations were intended for ananimal pharmacokinetics (PK) study. The capsules were filled manuallyusing filling funnel/stand and tamper into capsules size #9. Capsuleswith 100 mg Fenretinide were filled manually in capsules size #1 exceptlots 005a to 005c and 007a to 007c which were encapsulated in 00capsules. Weight variations below 3% RSD were observed, so adequate drugcontent uniformity is expected.

Example 3: Analytical Testing

Fenretinide 100 mg prototype formulations were tested by assay, relatedsubstances and dissolution rates. The results obtained as well as samplepreparations are shown below.

According the results obtained for neat API, the total amount ofimpurities seems to be related to the preparations of the samples duringmanufacturing and analytical testing (Table 11). Different levels ofimpurities were observed for samples from a same lot of API depending onthe extraction solvent and for two lots of API with the same solvent.The lowest related substances were observed for API lot 1 using methanolas solvent.

TABLE 11 Impurities Observed for the API Sample API - Lot 1 API - Lot 1replicate Sample Water to dissolve capsule, Water to dissolve capsule,Water to dissolve capsule, preparation then complete to volume thencomplete to volume then complete to volume with methanol (MeOH) withacetonitrile (ACN) with acetonitrite (ACN) Final ratio Final ratio Finalratio 20% Water:80% MeOH 20% Water:80% ACN 20% Water:80% ACN RRT % AreaRRT % Area RRT % Area Related 0.22 0.01 0.25 0.01 0.25 0.01 Substances0.26 0.01 0.27 0.07 0.27 0.08 (% area) 0.28 0.07 0.72 0.18 0.49 0.020.49 0.02 0.90 0.14 0.72 0.19 0.72 0.18 0.93 0.29 0.90 0.20 0.93 0.05Total 0.70 0.93 0.40 Total 0.35 Total 0.89Formulation Lots L215-01001 to 005c

Assay and related substances results were obtained using two differentsample preparation methods.

The results obtained for the formulations 001 to 004 showed remarkabledifference depending on sample preparation. For all capsule formulationsthe better results were obtained using 20% Water: 80% ACN as solventsystem (Table 12). Moreover, formulations and API (Table 11) showedsimilar impurities profiles.

Assay of NMT 50% were obtained for all Gelucire® 44/14 containingformulations (lots 003 and 004 a, b and c). Using 20% Water: 80% ACN assolvent system the total amount of related substances was between 0.68and 0.85% (Tables 13 and 14).

For solid dispersion formulations (lots 005a, b and c (Table 17)) assayof about 98% and related substances of about 1% were obtained with 20%Water: 80% MeOH as extraction solvent. Preparations with 20% Water: 80%ACN were not tested.

TABLE 12 Analytical Testing Results for Formulations 001 and 002 SampleL215-01001 L215-01002 Dose strength 100 mg/capsule 100 mg/capsule SampleWater to dissolve Water to dissolve Water to dissolve Water to dissolvepreparation capsule, then capsule, then capsule, then capsule, thencomplete to volume complete to volume complete to volume complete tovolume with methanol with acetonitrile (ACN) with methanol withacetonitrile (ACN) Final ratio 20% Final ratio 20% Final ratio 20% Finalratio 20% Water:80% MeOH Water:80% ACN Water:80% MeOH Water:80% ACNAssay 87.9% 101.4% 44.1% 94.6% (n = 2: 83.4, 92.5) (n = 2: 101.3, 101.5)(n = 2: 46.7, 41.6) (n = 2: 94.3, 94.8) RRT % Area RRT % Area RRT % AreaRRT % Area Related 0.22 0.01 0.25 0.01 0.26 0.02 0.25 0.01 substances0.26 0.01 0.27 0.07 0.28 0.13 0.27 0.08 (% area) 0.28 0.08 0.72 0.180.49 0.03 0.71 0.18 0.49 0.02 0.90 0.14 0.72 0.30 0.90 0.13 0.72 0.190.93 0.29 0.90 0.32 0.93 0.27 0.90 0.22 Total 0.70 0.93 1.02 Total 0.670.93 0.68 Total 1.81 Total 1.20

TABLE 13 Analytical Testing Results for Formulation 003 SampleL215-01003 Dose strength 100 mg/capsule Sample Water to dissolvecapsule, Water to dissolve capsule, preparation then complete to volumethen complete to volume with methanol with acetonitrile (ACN) Finalratio 20% Final ratio 20% Water:80% MeOH Water:80% ACN Assay 15.5% 50.2%(n = 2: 18.5, 12.5) (n = 2: 43.6, 56.8) RRT % Area RRT % Area Related0.28 0.10 0.27 0.08 Substances 0.72 0.25 0.72 0.16 (% area) 0.90 0.650.90 0.20 0.93 2.18 0.93 0.41 Total 3.18 Total 0.85

TABLE 14 Analytical Testing Results for Formulation 004a SampleL215-01004a Dose strength 100 mg/capsule Sample Water to dissolvecapsule, Water to dissolve capsule, preparation then complete to volumethen complete to volume with methanol with acetonitrile (ACN) Finalratio 20% Final ratio 20% Water:80% MeOH Water:80% ACN Assay 13.0% 51.8%(n = 2: 12.7, 13.3) (n = 2: 59.9, 43.7) RRT % Area RRT % Area Related0.28 0.09 0.28 0.07 Substances 0.72 0.22 0.72 0.15 (% area) 0.90 0.560.90 0.15 0.93 1.85 0.93 0.31 Total 2.73 Total 0.68

TABLE 15 Analytical Testing Results for Formulation 004b SampleL215-01004b Dose strength 100 mg/capsule Sample Water to dissolvecapsule, Water to dissolve capsule, preparation then complete to volumethen complete to volume with methanol with acetonitrile (ACN) Finalratio 20% Final ratio 20% Water:80% MeOH Water:80% ACN Assay 11.0% 37.8%(n = 2: 10.4, 11.7) (n = 2: 35.5, 40.1) RRT % Area RRT % Area Related0.28 0.09 0.28 0.08 Substances 0.72 0.23 0.72 0.15 (% area) 0.90 0.760.90 0.16 0.93 2.41 0.93 0.36 Total 3.49 Total 0.74

TABLE 16 Analytical Testing Results for Formulation 004c SampleL215-01004c Dose strength 100 mg/capsule Sample Water to dissolvecapsule, Water to dissolve capsule, preparation then complete to volumethen complete to volume with methanol with acetonitrile (ACN) Finalratio 20% Final ratio 20% Water:80% MeOH Water:80% ACN Assay 14.4% 32.2%(n = 2: 14.8, 14.0) (n = 2: 32.0, 32.5) RRT % Area RRT % Area Related0.28 0.09 0.28 0.08 Substances 0.72 0.23 0.72 0.15 (% area) 0.90 0.500.90 0.16 0.93 1.65 0.93 0.36 Total 2.46 Total 0.75

TABLE 17 Analytical Testing Results for Formulation 005a to 005c SampleL215-01005a L215-01005b L215-01005c Dose strength 100 mg/capsule 100mg/capsule 100 mg/capsule Assay 97.9% 97.9% 98.3% (n = 2: 97.5, (n = 2:98.5, (n = 2: 96.8, 98.2) 97.4) 99.8) RRT % Area RRT % Area RRT % AreaRelated 0.27 0.07 0.22 0.01 0.27 0.07 Substances 0.28 0.12 0.27 0.050.28 0.13 (% area) 0.49 0.09 0.28 0.10 0.49 0.10 0.72 0.18 0.33 0.010.72 0.18 0.90 0.14 0.43 0.01 0.90 0.17 0.93 0.41 0.49 0.05 0.93 0.50Total 1.00 0.72 0.18 Total 1.14 0.90 0.14 0.93 0.40 Total 0.93 Samplepreparation Water to dissolve capsule, then complete to volume withmethanol Final ratio 20% Water:80% MeOHFormulation Lots L215-007a to 007c

SDI-DG formulations lots 007a to 007c were obtained by dry granulationof Fenretinide (40%)/Plasdone® K-29/32 (60%) spray-dried intermediate(SDI) material.

These sample preparations were done using water to dissolve the capsulesand then completing to volume with acetonitrile for a final ratio 20%Water:80% ACN. In some duplicate samples, the excipients formed a clumpin the sample solution which may account for the variability in thereplicate assay samples.

The results (Table 18) showed comparable assay values and total amountof related substances for all 3 formulations with increase in impuritiesof about 1% when compared to neat API (Table 11). Moreover, theseformulations showed different related substances profile when comparedwith neat API and SDI formulations lots 005a, 005b and 005c.

The dissolution testing was achieved using different dissolution media.In acid medium, no release was noted in 1 hour. At pH 6.8 between 67 and78% of the drug was released within 60 minutes. The dissolution ratedepended mostly on aqueous solubility of the filler (lactose or calciumhydrogen phosphate dehydrate) and not on presence of croscarmellose asdisintegrant.

TABLE 18 Analytical Testing Results for Formulation 007a to 007c SampleL215-01007a L215-01007b L215-01007c SDI (40% PVP) dry SDI (40% PVP) drySDI (40% PVP) dry granulation (slugging granulation (slugginggranulation (slugging method) tablet method) tablet formulation method)tablet formulation Croscarmellose + formulation Croscarmellose + Tab-80Emcompress Aerosil + Tab-80 Dose strength 100 mg/capsule 100 mg/capsule100 mg/capsule Assay 95.8% 97.2% 94.1& (n = 2: 98.9, 92.7) (n = 2:101.1, 93.2) (n = 2: 95.6, 92.5) RRT % Area RRT % Area RRT % AreaRelated 0.24 0.08 0.24 0.08 0.24 0.07 Substances 0.26 0.63 0.26 0.620.26 0.57 (% area) 0.28 0.29 0.28 0.29 0.28 0.27 0.49 0.54 0.49 0.590.49 0.52 0.72 0.19 0.72 0.19 0.72 0.19 0.90 0.06 0.90 0.06 0.90 0.060.93 0.14 0.93 0.15 0.93 0.14 Total 1.93 Total 1.97 Total 1.81 Time TimeTime (minutes) % Dissolved (minutes) % Dissolved (minutes) % DissolvedDissolution 10 5 10 1 10 0 Paddles, 100 20 5 20 1 20 0 rpm ramp to 30 430 0 30 0 200 rpm at 60 60 2 60 0 60 0 minutes 75 1 75 0 75 0 900 ml0.1N HCl + 2.0% SDS Dissolution 10 65 10 57 10 67 Paddles, 100 20 72 2065 20 77 rpm ramp to 30 75 30 65 30 78 200 rpm at 60 60 73 60 67 60 78minutes 75 75 75 65 75 81 900 ml pH 6.8 + 2.0% SDS Dissolution 10 66 1060 10 34 Paddles, 100 20 71 20 67 20 72 rpm ramp to 30 73 30 66 30 72200 rpm at 60 60 73 60 66 60 76 minutes 75 72 75 66 75 73 900 ml pH8.0 + 2.0% SDS Note: A stock standard diluted in the 80% acetonitrilewas used to quantitate the dissolution samples. The stock standard wasalso diluted in each of the dissolution medium, good recoveries wereobtained for these standard solutions.

Example 4: SDI—Crystal State of Formulation Lots L215-01005a,L215-01005b and L215-01005c

FIG. 1 shows that irrespective of the polymer used as co-precipitate,amorphous SDI was produced. Physical stability of the spray-driedamorphous solid dispersion formulations (spray-dried intermediate) wasevaluated by XRPD. Lots L215-01005a, 005b and 005c were tested aftermanufacturing (T=0) and after 3.5 months of storage at 5° C. (bulkpowder in amber glass bottles). As shown in FIG. 13, the amorphous formremained stable during storage period. The lot L215-01006b was testedafter 50 days under refrigerator conditions and as shown the drugremains in an amorphous state.

Example 5: Pharmacokinetics Study of Formulation Lots L215-01002,L215-01003, L215-01004b, L215-01004c, L215-01005b and L215-01005c inRats

The objective of this study was to determine pharmacokinetic profiles of6 Prototype formulations of Fenretinide, after an oral dosage in ratswith the aim to select the best prototype for further optimization.

Study Protocol

-   -   Animals: Sprague-Dawley, male rats, n=3 per each formulation    -   Dosage regimen: Oral capsule gavage, (2×5 mg)    -   PK Timepoints: 0, 0.5, 1, 2, 4, 7, 14 and 24 hours (0.5        ml/timepoint in K₂EDTA)

The studies were conducted at the Centre National de BiologieExperimentale (CNBE) of the Institut Nationale de Recherche Scientifique(INRS). Six different prototypes of fenretinide oral formulations wereprepared in number 9 cellulose capsules (6 capsules/formulation). Threerats per each formulation type (weight 326-365.7 g) were administered 2capsules each using special delivery syringe after overnight fasting.

Animal ID Product Group 1 Fenretinide 5 MG 1 2 Lot #: L215-01002PK 3MFG: 2013JN19 4 Fenretinide 5 MG 2 5 Lot #: L215-01003PK 6 MFG: 2013JN207 Fenretinide 5 MG 3 8 Lot #: L215-01004bPK 9 MFG: 2013JN20 10Fenretinide 5 MG 4 11 Lot #: L215-01004cPK 12 MFG: 2013JN20 13Fenretinide 5 MG 5 14 Lot #: L215-01005bPK 15 MFG: 2013JN21 16Fenretinide 5 MG 6 17 Lot #: L215-01005cPK 18 MFG: 2013JN21

Blood samples were taken by jugular vein (0.5 ml/sample) puncturesamples and the obtained plasma was analyzed (HPLC) by MsPharma Inc., inLaval. Details of the analytical method are as follows.

Analysis of Fenretinide content in plasma sample by HPLC-UV.

Principle:

The principle is to retain the compound using a C18 reverse phase and anHPLC with UV detection at 360 nm.

Definitions

MP=Mobile Phase; MeOH=Methanol; HCl=Hydrochloric acid; RT=Roomtemperature; % RSD=% of relative standard deviation; % RD=% of relativedeviation

C or [ ]=concentration in pg/ml, ws=sample weight, V=Volume of samplepreparation, Ae=response of the sample, As=response of the standard,D=dilution factor, S/N=Signal-to-Noise ratio,

4-HPR=Fenretinide; 4-MPR=N-(4-methoxyphenyl)retinamide.

Materials:

HPLC with a C18 Inertsil ODS-3, 250×4.6 mm, 5 μm

MeOH, Water, syringe filter PVDF 0.45 μm, 4-HPR and 4-MPR.

Procedure:

The HPLC Conditions for plasma samples analysis are:

Column: C18 Inertsil ODS-3, 5 μm, 4.6 × 250 mm or equivalent Wavelength:360 nm Flow rate*: 1.0 mL/min. Temperature: 35° C. Injection Volume: 100μL Time % % MP (Gradient): (min) Water MeOH 0 75 25 2 75 25 3 1 99 14 199 14.01 75 25 18 75 25

The concentration of each sample was determined using the calibrationcurve forced through zero of the area ratio from the internal standardpeak (N-(4-ethoxyphenyl)retinamide, 4-EPR) and the concentration(μg/mL).

${{Calculation}\mspace{14mu}{{\mu g}/{mL}}} = {{\frac{Ae}{m}\mspace{14mu}{Where}\mspace{14mu} m} = {Slope}}$Results

The results are illustrated in FIG. 2 and in the Tables 19 and 20 below.The obtained plasma concentrations indicate relatively slow absorptionof Fenretinide (appearing at 1-2 hrs post-dose). T_(max) between 4 and 7hours was observed for all types of formulations tested, indicating thatthe absorption occurs probably mainly in the lower small intestine. InGroups 1-4, the C_(max) (14.4-21 ng/mL) and AUC_((0-24h)) (86-196ng-h/mL) were lower compared to the groups 5 and 6. The eliminationhalf-life was determined only for Groups 5 and 6 and was consistent withthe literature data for fenretinide (7-10 hrs) in rats. The highestfenretinide plasma exposure was observed in Group 5 (4188.7 ng-h/mL),followed by group 6 (3146.2 ng-h/mL).

TABLE 19 Summary of mean Fenretinide plasma concentration Time (h) Gr 1Gr 2 Gr 3 Gr 4 Gr 5 Gr 6  0    0.000  0.000  0.000  0.000   0.000  0.000  0.5  0.000  0.000  0.000  0.000   0.000   0.000  1    0.000 0.000  3.850  8.523   4.017   4.383  2    3.597  7.903  3.923  4.317 18.563  24.060  4   20.137 14.350 15.657 10.437 367.857 297.517  7  21.063 13.477  7.710 15.017 468.057 217.337 14    4.177  0.000  0.000 0.000  81.070 123.060 24    0.000  0.000  0.000  0.000  41.760  46.077Clinical Observations:

-   -   Gr. 2—⅓ rats did not received complete dose (second capsule        partially delivered)    -   All groups—increased water consumption observed in all rats        after 2 hrs post-dose

TABLE 20 PK parameters after single oral dose of Fenretinide in ratsParameter (Units) Group 1 Group 2 Group 3 Group 4 Group 5 Group 6C_(max) (ng/mL)  21.1  14.4 15.7  15.0  468.1        297.5     T_(max)(h)^(a)   7     4    4     7      7            4       t_(1/2e(h)) nd nd nd nd   10.4447131    7.053131 VD (L) nd nd nd nd  89.7         81.1      AUC₀₋₂₄ (ng·h/mL) 196.6 115.1 86.5 114.04188.7       3146.2     

Example 6: Pharmacokinetics Study of Formulation Lots L215-01005a,L215-01005c, L215-01007a, L215-01007b and L215-01007c in Rats

The objectives of this study were to continue the optimization of themost promising prototypes studied in the previous pharmacokinetic studyin rats. A new series of optimized prototypes was produced (L215-01007a,L215-01007b and L215-01007c) and compared with some of the previousprototypes (L215-01005a and L215-01005c) and with the originalFenretinide corn-oil formulation. The formulation with the mostfavourable pharmacokinetic profile for development of formulationsuitable for treatment of human subjects was selected for futurestudies.

Study Protocol

-   -   Animals: Sprague-Dawley, female rats, n=3 per each formulation    -   Dosage regimen: Single dose by oral gavage    -   PK Timepoints: 0, 1, 2, 4, 5, 7, 14, 24 and 48 hours (0.5        ml/timepoint)

The studies were conducted at the Centre National de BiologieExperimentale (CNBE) of the Institut Nationale de Recherche Scientifique(INRS). Six different fenretinide oral formulations were studied; 5solid formulations presented in number 9 gelatine capsules (min 6capsules/formulation, Table 21) and one liquid suspension formulation incorn oil.

TABLE 21 Overview of the study groups Mean Ani- Dose/ mal Dose Group IDFormulation Group Dose/rat (mg/rat) (mg/kg) 20 Fenretinide 7 300 μL 11.432.0 21 38.02 mg/mL 300 μL 11.4 22 Corn oil formulation 300 μL 11.4 23Fenretinide 8 2 capsules 5 13.3 24 2.5 mg/capsule 2 capsules 5 25 Lot #:L215-01007a 2 capsules 5 Mfg: 2013JL23 26 Fenretinide 9 2 capsules 513.5 27 2.5 mg/capsule 2 capsules 5 28 Lot #: L215-01007b 2 capsules 5Mfg: 2013JL23 29 Fenretinide 10 2 capsules 5 13.8 30 2.5 mg/capsule 2capsules 5 31 Lot #: L215-01007c 2 capsules 5 Mfg: 2013JL23 32Fenretinide 11 2 capsules 10 27.4 33 5 mg/capsule 2 capsules 10 34 Lot#: L215-01005a 2 capsules 10 Mfg: 2013JL16 35 Fenretinide 12 2 capsules10 26.8 36 5 mg/capsule 2 capsules 10 37 Lot #: L215-01005c 2 capsules10 Mfg: 2013JL22

The suspension formulation was prepared by extraction of the content of3 McNeil soft capsules containing each 100 mg of 4-HPR (obtained fromthe National Cancer Institute, National Institutes of Health, Bethesda,Md., USA). The content of the three capsules was mixed, diluted toobtain a final concentration between 35-40 mg/ml by adding corn oilfollowed by vigorous mixing for 4 minutes. The concentration of 4-HPR inthe final corn-oil formulation for animal dosing was confirmed by HPLCanalysis (MsPharma Inc.) prior to gavaging animals, and contained 38.02mg/mL of 4-HPR.

Three female Sprague-Dawley rats per group (weight 337.1-390.3 g)received 2 capsules each per os using special capsule delivery syringe.Groups 8, 9 and 10 received the optimized formulations (L215-01007a,L215-01007b and L215-01007c) and Groups 11 and 12 the prototypeformulations (L215-01005a and L215-01005c). The corn oil basedformulation was administered to 3 rats of Group 7 in a volume of 300 μLper rat using a stainless steel gavage needle. Following the dosing ofthe drug formulations there were no clinical observations in any of thegroups up to 48 hours, the last time point studied, at which the animalswere euthanized.

Blood samples were collected into K2EDTA tubes at predeterminedtimepoints by jugular vein puncture (˜0.5 mL/sample) alternating theleft and right jugular veins sites. The blood samples were kept on wetice protected from direct light until centrifugation. The obtainedplasma was immediately transferred to amber Eppendorf tubes and storedat −20° C. until analysis. The animals were fasted overnight before thedosing (water ad libitum) and were fed approximately 2 hours after thedosing.

The obtained plasma was analyzed by HPLC at MsPharma Inc. (Laval, QC)for content of 4-HPR and 4-MPR, the main metabolite of 4-HPR, using thebioanalytical method described above. After integration of the peakareas, data were exported to an Excel® (Microsoft®) spreadsheet. TheExcel spreadsheet was used for calculation of 4-HPR and 4-MPRconcentrations in rat plasma and for descriptive statistics. Meanconcentration-time data for each timepoint were analyzed using PCSolution Software 2.0™ (Summit Research Services, Montrose, Colo., USA)with nominal sampling times. C_(max) and T_(max) were confirmed byinspection of observed data. Areas under the plasma meanconcentration-time curves (AUC) were estimated using the lineartrapezoidal rule and reported as AUC₍₀₋₄₈₎. Apparent terminal half-life(t_(1/2)) was determined, when possible, by linear regression analysisof three concentrations that appeared to be on the terminal eliminationphase of the mean concentration time-curve. The bioanalytical data andresults for 4-HPR and 4-MPR are presented in FIGS. 3-5 and in Tables22-27.

TABLE 22 Summary of mean 4-HPR plasma concentration (ng/mL, n = 3) Group7 Group 8 Group 9 Group 10 Group 11 Group 12 Corn oil OptimizedOptimized Optimized Prototype Prototype Formulation L215-01007aL215-01007b L215-01007c L215-01005a L215-01005c Time (h) Mean SEM MeanSEM Mean SEM Mean SEM Mean SEM Mean SEM  0   0.0  0.0   0.0  0.0   0.0 0.0   0.0  0.0   0.0  0.0   0.0  0.0  1  38.6  0.6  15.3 14.4  57.744.9   2.0  1.2   7.9  4.0  12.3 12.3  2  99.5 12.9  60.3 40.8 126.163.8  43.1 19.1  21.9  9.7  54.8 31.1  4 181.7 22.3 174.1 40.2 279.344.8 171.5 82.4  93.2 33.7 223.9 24.6  5 172.6 23.5 215.9 36.7 317.428.6 172.1 89.5 149.4 14.8 166.7 32.3  7 200.3 40.0 310.8  1.9 340.077.0 212.3 57.7 156.1 48.7 132.1 15.9 14  58.8 19.4  63.3 11.6  76.6 9.3  57.9 12.8  31.4  9.4  48.0 12.9 24  12.3  6.3  22.0  4.0  34.3 1.5  22.6  3.8  17.0  4.1  11.3  0.8 48   4.6  1.3   3.9  0.6   5.9 1.0   4.1  1.1   2.8  1.4   3.3  0.9

TABLE 23 Calculated PK parameters after single oral dose of 4-HPR inrats Group 7 Group 8 Group 9 Group 10 Group 11 Group 12 Corn oilOptimized Optimized Optimized Prototype Prototype Parameter (Units)Formulation L215-01007a L215-01007b L215-01007c L215-01005a L215-01005cMean Dose (mg/kg)   32.0   13.3   13.5   13.8   27.4   26.8 C_(max)(ng/mL)  200.3  310.8  340.0  212.3  156.1  223.9 T_(max)(h)^(a)    7     7      7      7      7      4   AUC₀₋₄₈ (ng·h/mL) 2385.1 3047.43977.4 2464.1 1696.3 1915.2 t_(1/2e(h))   10.1    8.6    9.3    9.1   9.7    9.5 VD (L)   69.8   20.4   16.8   26.5   82.4   71.2 Clearance(mL/h) 4243.0 1640.8 1257.1 2029.2 5895.3 5221.3

TABLE 24 4-HPR plasma exposure normalized to 20 mg/kg oral dose Group 7Group 8 Group 9 Group 10 Group 11 Group 12 Corn oil Optimized OptimizedOptimized Prototype Prototype Parameter (Units) Formulation L215-01007aL215-01007b L215-01007c L215-01005a L215-01005c AUC₀₋₄₈ (ng·h/mL)1490.68 4584.87 5911.44 3579.49 1238.96 1429.15 C_(max) (ng/mL)  125.18 467.54  505.30  308.36  114.04  167.07 % of Corn oil AUC  100    307.6   396.6   240.1    83.1    95.9 

TABLE 25 Summary of mean 4-MPR concentration after 4-HPR oral dose(ng/mL, n = 3) Group 7 Group 8 Group 9 Group 10 Group 11 Group 12 Cornoil Optimized Optimized Optimized Prototype Prototype FormulationL215-01007a L215-01007b L215-01007c L215-01005a L215-01005c Time (h)Mean SEM Mean SEM Mean SEM Mean SEM Mean SEM Mean SEM  0 0.0 0.0  0.00.0  0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0  1 0.0 0.0  0.0 0.0  0.0 0.0 0.00.0 0.0 0.0 0.0 0.0  2 1.9 1.0  0.9 0.9  3.2 2.3 0.0 0.0 0.0 0.0 0.5 0.5 4 7.3 0.9  5.7 2.8  9.7 2.9 4.7 3.0 1.1 1.1 9.6 2.0  5 7.3 1.2  7.5 1.112.9 2.2 6.5 4.3 3.9 0.2 6.2 1.5  7 9.6 1.7 13.9 0.7 15.7 3.5 9.5 4.06.6 2.3 7.6 1.4 14 5.3 1.5 11.2 2.0 13.7 4.2 5.4 1.0 3.1 1.6 2.9 1.7 240.0 0.0  1.6 0.8  3.8 0.3 0.8 0.8 0.9 0.9 0.0 0.0 48 0.0 0.0  0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0

TABLE 26 Calculated PK parameters of 4-MPR after single oral dose of4-HPR in rats Group 7 Group 8 Group 9 Group 10 Group 11 Group 12 Cornoil Optimized Optimized Optimized Prototype Prototype Parameter (Units)Formulation L215-01007a L215-01007b L215-01007c L215-01005a L215-01005cC_(max) (ng/mL)   9.6  13.9  15.7   9.5  6.6  9.6 T_(max)(h)^(a)   7    7     7     7    7    4   AUC₀₋₂₄ (ng·h/mL) 112.5 207.2 289.9 119.978.5 83.1 % of fenretinide AUC   4.7   6.8   7.3   4.9 4.6  4.3

TABLE 27 4-MPR plasma exposure normalized to 20 mg/kg oral dose of 4-HPRGroup 7 Group 8 Group 9 Group 10 Group 11 Group 12 Corn oil OptimizedOptimized Optimized Prototype Prototype Parameter (Units) FormulationL215-01007a L215-01007b L215-01007c L215-01005a L215-01005c C_(max)(ng/mL)   5.98  20.96  23.40  13.82  4.85  7.14 AUC₀₋₄₈ (ng·h/mL)  70.41311.69 430.81 174.20 57.33 62.02 % of Corn oil AUC 100    442.7  611.9 247.4  81.4  88.1 

The 4-HPR plasma concentration profile of all the formulations showed asimilar pattern (FIG. 3); very low plasma levels present at 1 hour afterdose and maximum concentrations of 156.1-340 ng/mL (C_(max)) reaching at7 hours (T_(max)), (Tables 22 and 23) indicated a relatively slowabsorption occurring probably in the lower small intestine. 4-HPRappeared to be highly distributed in bodily tissues. The eliminationfollowed a first order kinetic; the half-life (t_(1/2)) was between 8.6h and 10.1 h. Plasma exposure AUC_((0-48h)) varied significantly amongthe different formulation types (1696.3-3977.4 ng·h/mL). 4-HPR exposuredata normalized to 20 mg/kg dose (Table 24) indicate relatively highexposure achieved with all 3 optimized formulations (Groups 8, 9 and 10)2.4-4 times higher when compared to the 4-HPR corn oil formulation(1490.7 ng·h/mL) while the prototype formulations (Groups 11 and 12) ledto an exposure similar to the corn oil formula. The highest fenretinideplasma exposure was observed in Group 9 (5911.4 ng·h/mL).

Statistical analysis of raw plasma concentrations of the three optimizedformulations using Two-way ANOVA analysis followed by All PairwiseMultiple Comparison Procedures (Student-Newman-Keuls Method)demonstrated that the difference in the mean values among the differentlevels of group and time points were greater than would be expected bychance (P=0.004). There was no statistical difference in plasmaconcentrations among the 3 groups of animals receiving optimizedformulation at all but one time points tested. Only at the 5 hr timepoint there was statistically significant (p=0.036) difference betweenGroups 9 and 10.

The plasma profile of 4-MPR (main metabolite of 4-HPR) generallyfollowed the profile of 4-HPR (FIG. 4). 4-MPR appeared in plasma at 2-4hours after the dosing, reached relatively low C_(max) (9.5-15.7 ng/mL)at T_(max) of 4-7 hours (Tables 25 and 26). Plasma exposure of 4-MPRrelative to the parent 4-HPR (FIG. 5) varied between 4.3% (Gr. 12) and7.3% (Gr. 10). 4-MPR exposure data normalized to 20 mg/kg of 4-HPR dose(Table 27) indicated 2.5-6.1 times higher 4-MPR levels in the optimizedformulations when compared to the corn oil formulation, following anapproximately equivalent increase in 4-HPR plasma exposure.

The above data shows that compared to the corn oil formulation, allthree optimized formulations lead to higher plasma exposure of 4-HPR(2.4-4 times) after a single oral dose in the rat model. Highestapparent exposure was achieved with the formulation dosed in Group 9(Lot #: L215-01007b).

Example 7: Solid Dispersion Optimization

Solvent System Optimization, Solid Loading Maximization and Scale-Up

Based on the results of the pharmacokinetics studies in animalsdescribed above, the solid dispersion approach (lots L215-01005 toL215-01008) was selected for Fenretinide clinical trial material (CTM)manufacturing.

The first step of the optimization was to replace the methanol byethanol to avoid using multiple solvents with higher toxic potential.Indeed, methanol is a class 2 solvent (solvent to be limited) inpharmaceutical products with a concentration limit of 3000 ppm whileethanol is a class 3 solvent (solvent with low toxic potential) with aconcentration limit of 5000 ppm (U.S. Department of Health and HumanServices, Food and Drug Administration, Center for Drug Evaluation andResearch (CDER), Guidance for Industry Q3C Impurities: ResidualSolvents, December 1997). For information, Dichloromethane (DCM) is alsoa class 2 solvent with a concentration limit of 600 ppm.

When solubilised with PVP K30 in a ratio of 40/60 w/w, it was determinedthat the solubility of Fenretinide in ethanol 100% was about 34 mg/mland over 50 mg/ml in DCM 100% or a mixture of ethanol/DCM 50/50 v/v. Thesystems with ethanol had similar viscosity value and higher than thesystem composed of DCM only (Table 28) suggesting a higher solid loadinglimit with increasing amount of DCM.

TABLE 28 Viscosity at RT of Fenretinide/PVP K30 40/60% w/w Solution inVarious Solvent Systems Solvent System Viscosity (Fenretinideconcentration) (mPas) EtOH 100% (34 mg/ml) 6.45 EtOH/DCM 50/50 v/v (40mg/ml) 6.00 DCM 100% (40 mg/ml) 2.95

At lower solid loading (7.5% w/w), the influence of DCM on the viscosityof the solution was lower as shown by lots L215-01009A, B and C (Table29). For these lots, it was found that the yield of the spray-dryingprocess increases with increasing ratio of DCM in solution. Asignificant amount of volatile compounds (between 3.13 and 4.24%) waspresent in the SDI (FIG. 9). GC analysis showed that in all samples,ethanol and DCM was among the volatile compounds (Table 29). Theremaining volatile compound was probably water. The residual amount ofDCM in the SDI varied also with the ethanol/DCM ratio, a higher ratio ofDCM in the solution resulted in higher amounts of DCM in the SDI. Highlevel of ethanol was found in all SDI of lot L215-01009 (Table 29). Thehigher amount of ethanol found in the lots L215-01009B and C compared tothe lot L215-01009A probably resulted from the lower inlet temperatureused to produce the SDI lots L215-01009B and C. FIGS. 6B-D show that theDCM/ethanol ratio had a limited influence on the SDI particlemorphology. Most of the particles were irregular shapes and appeared ascollapsed sphere, but slightly more spherical particles with smoothsurface was observed within the SDI of lot L215-01009C. The DCM/ethanolratio did not influenced the crystal state of the SDI, where for thelots L215-01009A, B and C amorphous material was produced (FIG. 7). Nothermal event, particularly no glass transition, before 150° C. wasidentified on the DSC thermogram of the SDI lots L215-01009A, B and Csuggesting a stability of the amorphous state at under normaltemperature condition used for dry granulation and encapsulation process(FIG. 8). According to these results, it was selected to use DCM onlyfor the spray dry solution for further development.

With DCM only as solvent, it was possible to increase the solid loadingup to 20%. At this solid loading, the viscosity of the solution of lotL215-01010 was high (Table 29) and more difficult to spray. Theatomization of the solution was incomplete causing sticking of materialon the inside wall of the drying chamber and filament like particle(FIG. 6E). Despite these facts, the spray drying yield of the lotL215-01010 was comparable to the lot L215-01009C. The SDI of lotL215-01010 was also in an amorphous state (FIG. 7), no thermal eventbefore 150° C. was observed on its DSC thermogram (FIG. 8) and about2.5% of volatile compounds was release from the SDI when heated up to125° C. (FIG. 9).

To improve the SDI particle morphology, the solid loading in thesolution was decreased to 12.5% for the lot L215-01011. FIG. 6F showsthat at this solid loading, particle with morphology similar to that oflot L215-01009 was obtained. As previously, the SDI of the lotL215-01011 was in an amorphous state (FIG. 7), and had similar thermalproperties to the lots L215-01009 and L215-01010 (FIGS. 8 and 9). Withincrease batch size, 250 g, the maximum spray drying yield among thespray drying trials was achieved with the lot L215-01011 (Table 29). Theincrease batch size was also associated with an increase of the residualamount of dichloromethane in the SDI (Table 29), which was easilyremoved by the secondary drying at RT and under −15 inHg vacuum whereafter 16 hours, the totality of the DCM was almost removed (Table 29).The secondary drying did not modify the amorphous sate of the SDI (FIG.7). The SDI lot L215-01011 was characterized by an assay value of 96.3%and with a low level of related substance (0.43%) (Table 31). Thecomposition of the lot L215-01011 and the process parameters used toproduce it are thus recommended for CTM manufacturing of Fenretinide.

TABLE 29 Fenretinide/PVP K30 40/60% w/w SDI Process Related Data SprayResidual Viscosity Drying Solvent Lot (mPas) Yield (%) (ppm) L215-01009A3.72 72.3 EtOH: 3 687 DCM: 63 L215-01009B 3.43 79.0 EtOH: 5 805 DCM: 500L215-01009C 2.88 86.0 EtOH: 4 586 DCM: 2 539 L215-01010 10.2 89.6 DCM: 8619 L215-01011 3.42 95.3 DCM: 13 616 DCM: 19* *After secondary drying.SDI Direct Encapsulation

Uniform final blend was obtained for lots L215-01012A and L215-01012B.The encapsulation of both lots involved to force with a moderatepressure the final blend inside the capsule bodies with stainless steeltamping pin to reach the desired capsule weight. The capsule fill weightof lot L215-01012B was slightly over the target weight (Table 30)resulting in assay value of 101% (Table 32). Compared to lot L215-01007(Table 19), the Fenretinide dissolution from the capsule of lotL215-01012B in pH 6.8+2% SDS (Table 33 and FIG. 10) was slower buthigher plateau value was reached. These results indicated that a drygranulation step is not necessary to produce Fenretinide 100 mg capsulebased on Fenretinide/PVP K30 40/60% w/w SDI. To reduce stress on theSDI, simplify process and maximize process throughput, directencapsulation is thus preferred for CTM manufacturing.

TABLE 30 Fenretinide 100 mg HGC Direct Encapsulation Formulation lotL215-01012B (n = 100) and Fenretinide Placebo Formulation lotL215-01013P (n = 16) Fill Weight Statistics Fill weight Stdev RSD Min.Max. Lot Ave. (mg) (mg) (%) (mg) (mg) L215-01012B 463.5 6.0 1.3 445.9475.8 L215-01013P 449.2 4.5 1.0 436.9 455.9Short Term Stability Study

Tables 34 and 35 present the appearance, moisture, assay and totalrelated substance in SDI lot L215-01011 and Fenretinide 100 mg HCG lotL215-01012B incubated with closed cap at 5° C., 25° C./60% RH and 40°C./75% RH after 1 month and 9 months. The dissolution profile of lotL215-01012B is presented in Table 33 and in FIG. 10.

After an incubation of 1 month no modification of the sample'sappearance was noticed. At each time point, the moisture content wassimilar in all samples irrespectively of the incubation condition andremained under 5%, suggesting that the addition of moisture protectionwith the product may not be necessary when stored in a sealed HDPEbottle. Stability of amorphous forms was maintained for 1 month underall 3 conditions of temperature and humidity and remained stable evenafter 9 months of incubation at 5° C. (FIG. 14).

Non-refrigerated samples (25° C./60% RH and 40° C./75% RH) showed adecrease of the assay value associated with the increase of the amountof related substance over time. For samples stored at 5° C., assay wasof 96%, remaining as previous time-point and the total amount of relatedsubstances increased by 0.24%. However, further degradation duringsubsequent storage (9 months) occurred with a total amount of relatedsubstances of 8.36% area and largest impurity 2.85% area.

FIGS. 11 and 12 show that the amorphous form of Fenretinide remainedstable even after a 1-month incubation at 40° C./75% RH. As shown by theslope of the dissolution profiles of the L215-01012B samples (FIG. 10),the dissolution rate of Fenretinide did not appear to be influenced bythe decreased of the assay value suggesting a certain level ofrobustness for the direct encapsulation approach.

TABLE 31 Assay and Related Substances for SDI lot L215-01011 Incubatedat 5° C., 25° C./60% RH and 40° C./75% RH Sample L215-01011 SDIFenretinide/PVP K30 Dose strength 40% Drug load 5° C. 25° C./60% RH 40°C./75% RH Appearance T = 10 Yellow fine powder Yellow fine powder Yellowfine powder days T = 1 Yellow powder Yellow powder Yellow powder month T= 9 Yellow powder N/A N/A months 5° C. 25° C./60% RH 40° C./75% RHMoisture (KF) T = 10 2.7% 2.7% 2.9% days T = 1 3.2% 3.0% 3.5% month T =9 2.9% ND ND months Assay T = 0 96.3% (n = 2: 94.0, 98.7) 5° C. 25°C./60% RH 40° C./75% RH T = 10 96.2% 93.1% 73.2% days (n = 2: 97.2,95.3) (n = 2: 93.2, 93.1) (n = 2: 72.9, 73.4) T = 1 96.0% 77.4% 54.1%month (n = 2: 96.4, 95.7) (n = 2: 77.3, 77.6) (n = 2: 54.2, 53.9) T = 972.7% N/A N/A months (n = 2: 72.4, 73.0) Related T = 0 Total: 0.43% areaSubstances (% area) Largest impurity: 0.15% area @ RRT 0.72 5° C. 25°C./60% RH 40° C./75% RH T = 10 Total: 0.95% area Total: 1.52% areaTotal: 7.68% area days Largest impurity: Largest impurity: Largestimpurity: 0.37% area @ 0.55% area @ 2.39% area @ RRT 0.22 RRT 0.27 RRT0.27 T = 1 Total: 1.19% area Total: 7.35% area Total: 17.92% area monthLargest impurity: Largest impurity: Largest impurity: 0.38% area @ 2.40%area @ 4.69% area @ RRT 0.27 RRT 0.27 RRT 0.27 T = 9 Total: 8.36% areaN/A N/A months Largest impurity: 2.85% area @ RRT 0.27 RRT = relativeresponse time

TABLE 32 Assay and Related Substances for Fenretinide 100 mg HGC lotL215-01012B Incubated at 5° C., 25° C./60% RH and 40° C./75% RH ClosedCap for 1 Month Sample L215-01012B SDI (40% PVP dry granulationCroscarmellose + Emcompress ® dehydrate (size 00 Capsule) Dose strength100 mg/capsule 5° C. 25° C./60% RH 40° C./75% RH Appearance T = 10 Twopiece orange Two piece orange Two piece orange days capsules capsulescapsules T = 1 Yellow powder inside Yellow powder inside Yellow powderinside month orange capsule orange capsule orange capsule 5° C. 25°C./60% RH 40° C./75% RH Moisture (KF) T = 10 3.1% 3.3% 3.5% days T = 13.75  4.1% 4.7% month Assay T = 0 101.0% (n = 2: 101.5, 100.6) 5° C. 25°C./60% RH 40° C./75% RH T = 10 98.5% 94.5% 92.1% days (n = 2: 98.1,98.9) (n = 2: 93.3, 95.8) (n = 2: 92.0, 92.2) T = 1 96.6% 87.5% 83.7%month (n = 2: 96.7, 96.5) (n = 2: 87.5, 87.5) (n = 2: 83.7, 83.6)Related T = 0 Total: 0.40% area Substances (% area) Largest impurity:0.18% area @ RRT 0.72 5° C. 25° C./60% RH 40° C./75% RH T = 10 Total:0.84% area Total: 1.90% area Total: 2.30% area days Largest impurity:Largest impurity: Largest impurity: 0.19% area @ 0.54% area @ 0.68% area@ RRT 0.72 RRT 0.50 RRT 0.49 T = 1 Total: 1.79% area Total: 5.63% areaTotal: 6.91% area month Largest impurity: Largest impurity: Largestimpurity: 0.41% area @ 1.43% area @ 1.81% area @ RRT 0.27 RRT 0.27 RRT0.49 RRT = relative response time

TABLE 33 Dissolution for lot L215-01012B Incuvated at 5° C., 25° C./60%RH and 40° C./75% RH Close Cap for up to 1 Month Sample L215-01012B SDI(40% PVP) dry granulation Croscarmellose + Emcompress dehydrate (size 00Capsule) Dose strength 100 mg/capsule Time % (minutes) DissolvedDissolution T = 0 10 22 Paddles, 100 rpm (n = 3) 20 42 ramp to 200 rpm30 55 at 60 minutes 60 83 900 ml 75 94 pH 6.8 + 2% SDS 5° C. 25° C./60%RH 40° C./75% RH Time % Time % Time % (minutes) Dissolved (minutes)Dissolved (minutes) Dissolved T = 10 10 21 10 15 10 15 days 20 44 20 3720 38 (n = 3) 30 56 30 53 30 54 60 78 60 84 60 82 75 90 75 94 75 90 T =1 10 18 10 13 10  8 month 20 44 20 30 20 34 (n = 2) 30 60 30 44 30 52 6084 60 71 60 75 75 91 75 80 75 81

Example 8: Further Optimization of the Solid Fenretinide Dispersion

Table 34 provides a description of the materials used in this study.

TABLE 34 Material (Commercial Name) Lot # Supplier Fenretinide C00324Cedarburg Povidone (Plasdone K-29/32) C00450 ISP PVPK30 Povidone(Plasdone K-12) 0001596798 Ashland PVPK12 Hydroxypropyl methylcellulose2103280 Shin-Etsu acetate succinate (HPMCAS) Butylated hydroxyanisole(BHA) C00473 A&C Butylated hydroxytoluene (BHT) C00474 A&CDichloromethane HPLC grade 53130 EMD Methanol HPLC grade 54702 EMDMicrocrystalline cellulose C00537 FMC (Avicel PH-102) Dibasic CalciumPhosphate C00084 JRS Dihydrate (Emcompress) Croscarmellose Sodium C00020Blanver (Solutab Type A) Acid ascorbic 20747096 A&C Magnesium StearateVegetal C00124 Peter Greven grade MF-2-V Opadry AMB II 88A180040 whiteWP740303 Colorcon Empty Hard Gelatin Capsules, C00159 Capsugel Size 00,CS, Orange Opaque

Solid dispersions or Spray-Dried Intermediates of Fenretinide wereobtained by spray drying technique (Tables 35 and 36).

Preparation of Solution:

The solutions were prepared by dissolving the powders in 400 ml of asolvent system (50:50% v/v of methanol/dichloromethane) at 5% of solids.The mixtures were stirred until all particles were dissolved.

Spray-Drying:

The solutions were processed using a Model GA32 Yamato™ Lab Spray Dryerwith the following operating parameters: internal nozzle diameter 711μm; between 15-20 ml/min feed rate; 65° C. inlet temperature; 35-40° C.outlet temperature; 1.5 kg/cm² atomization air, and 0.45 m³/min airflow. After spraying, the heating was stopped and the drying wascontinued for an additional 3 minutes at an outlet temperature less than45° C. The SDIs were collected in the receiving flask (after cyclone)for yields of 62-71%. Also, the samples were protected from light duringall steps of the formulation development and stored at −20° C. untiluse.

TABLE 35 With/Without Antioxidants PVP and HPMCAS based Fenretinide SDIFormulations (20 g of solids/lot) Lot No. L215- L215- L215- L215- L215-Ingredients 01016 01017 01018 01019 01020 Fenretinide 100.0% 40.0% 40.0%40.0% 40.0% Povidone, type K-29/32 — 60.0% 59.8% — — HPMCAS — — — 60.0%59.8% BHA — —  0.1% —  0.1% BHT — —  0.1% —  0.1% MeOH-DCM (1:1 v/v) 400ml 400 ml 400 ml 400 ml 400 ml Total solid phase:  100%  100%  100% 100%  100% Lots L215-01021 and 022 are the placebos for the lotsL215-01018 and 020, respectively.

A new series of Fenretinide SDIs (Table 36) was prepared using increasedamount of antioxidants and two different grades of Povidone (PVPK30 andK12) as polymer. The modifications were based on improved stabilityresults obtained for lot L215-01018.

Preparation of Solution:

The solutions were prepared by dissolving the powders in 400 ml of asingle solvent (dichloromethane) at 5% of solids. The mixtures werestirred until all particles were dissolved.

Spray-Drying:

The solutions were processed using a Model GA32 Yamato™ Lab Spray Dryerwith the following operating parameters: internal nozzle diameter 711μm, about 10 ml/min feed rate; 60° C. inlet temperature; 31-40° C.outlet temperature; 1.5 kg/cm² atomization air, and 0.5 m³/min air flow.The SDI yield on receiving flask (after cyclone) were improved between76-83%.

Drug Products (DP)

Based on improved purity results, SDI lots L215-01023 and 027 werechosen to prepare Fenretinide 100 mg capsules and tablets (Tables 37 and38). Tablets were also coated with polyvinyl alcohol (PVA) basedmoisture barrier film coating at 10% weight gains (Opadry™ AMB II88A180040 white).

Final blends were obtained by dry granulation—slugging method. Firstpowders were sieved with a 30-mesh screen and mixed using a PK V-blenderfor 5 minutes at 25 RPM and 2 additional minutes after addition of thelubricant (magnesium stearate). The compacts (slugs) were produced usinga Carver single punch laboratory press with 12 mm die and punchescombination at 2-3 kP hardness. The granules were formed by crushing andpassing compacts through an 850 μm (20-mesh) screen.

475 mg dose was obtained by filling the granules into size 00 capsulesusing a Cooper filling capsule device. The tablets were compressed withGlobe Pharma™ Rotary Press using caplet shaped tooling 6.05×17.75 mm.Half quantity of the core tablets were coated with 10% weight gain ofOpadry™ AMB aqueous moisture barrier film coating system (20% solids)using an Aeromatic Strea™ fluid bed equipped with Wurster column andbottom spray nozzle system. The coating was carried out at inlettemperature 50-60° C., outlet temperature 45-50° C., spray rate 3-5g/min, atomization pressure 1.4-1.6 bars, Airflow 110-130 m³/h.

Characterization (XRPD, TGA, stability) was performed as describedabove.

Stability—crystalline API (lot C00324) and amorphous SDI. Thecrystalline API (lot C00324) and amorphous SDI powders, lots L215-01016to 020 were stored at 5° C., 25° C./60% RH and 40° C./75% RH in amberclosed and open bottles. The closed bottles were placed in double PEbags tightly closed and containing oxygen scavenger (StabilOx®) (1 unit)and desiccant (MiniPax® Sorbent Packets, 2 units), followed by aluminumseal.

Stability—amorphous SDI vs. Drug products. SDI powders (about 0.6 gram),lots L215-01023 to 027 and 100 mg Fenretinide drug product formulationslots L215-01028 to 033 were stored at 5° C. and 25° C./60% RH in UlinePoly bags closed and containing desiccant (MiniPax® Sorbent Packets, 2units), followed by aluminum seal (2× Statshield® moisture barrier bagswith Nitrogen purge).

Fenretinide, PVP, BHA and BHT are all very soluble in a mixture ofmethanol/dichloromethane (1:1) as well as in pure dichloromethane. Asresults, clear solutions of API (lot L215-01016) as well as API/PVPsamples (lot L215-01017) and API/PVP/BHA/BHT (L215-01018 and 023 to 027)were obtained rapidly. Formulations containing HPMCAS resulted in aturbid slightly viscous liquid. Irrespective of the formulation,amorphous form of Fenretinide was produced by spray drying for all lotswith a typical amorphous halo XRPD diffraction pattern (FIGS. 15 and16).

The amount of volatile components (residual dichloromethane and methanolfrom spray-drying process) was determined by TGA. Amorphous API (lotL215-01016) and API/HPMCAS SDIs (lots L215-01019 and 020) showed lessthan 1% of volatile components content. PVP containing SDIs (lotsL215-01017 and 018) showed an increase at 4.2% of volatile content thatcould be a result of the water content or residual solvent affinity withPVP. Identification and quantification of volatile compounds could beobtained by gas chromatography. For spray drying of lots L215-01023 to027, pure dichloromethane was used as a solvent, the drying temperatureof 60-65° C., spray rate of about 10 ml/min and drying time afterspraying was between 5-7 minutes. Under these conditions TGA showed amass loss of 1.77-3.65% between RT and 100° C. lower than previous PVPcontaining SDIs (lots L215-01017 and 018) prepared withdichloromethane/methanol solvents.

Stability #1 (API vs SDI)—Lots L215-01016 to 020

SDIs were tested for assay and related substances and compared withamorphous and crystalline API. Initial results obtained are shown inTable 39. The amorphous API (lot L215-01016) showed lower assay andincreased amount of related substances when compared with un-processedcrystalline form (99.2% and 0.25%, respectively). However, when using astabilizing polymer, the amount of related substances for SDIs lotsL215-01017 and L215-01019 were lower when compared to the pure APIamorphous form. The addition of antioxidants within the SDIs furtherincreased stability (L215-01018 and L215-01020). Stability wasinvestigated after 1 and 3 months under 5° C., 25° C./60% RH and 40°C./75% RH (open and closed cap glass amber bottles). The results arealso shown in Table 39. Antioxidants appear to prevent or retarddegradation for samples stored under 5° C. and 25° C./60% RH open cap.SDI lots L215-01017 and L215-01018 based on Plasdone™ showed lessdegradation than the SDI lots using HPMCAS.

Stability #2 (SDI vs DP)—Lots L215-01023 to 033

Five new Fenretinide SDI formulations were produced based on most stablelot L215-01018 (API, PVP as polymer and BHA+BHT as antioxidants) butusing dichloromethane as single solvent. Drug loading ranging from 30 to50% and the use of 2 grades of Plasdone™ K12 and K30 were evaluated.Initial testing results (Table 40) did not show major differencesbetween SDIs and also compared with the raw crystalline API. However,lots L215-01023 and 026 showed the highest assay values and yields andlowest water content and total related substances. These resultsconfirmed the suitability of the 40% drug loading using both PVP K30 andPVP K12 along with BHA and BHT as antioxidants.

Stability of lots L215-01023 to 27 was investigated after 1 and 3 monthsunder 5° C. and 25° C./60% RH, and at 6 months under 5° C., in closedcontainers. The results are shown in Table 41. Stability of formulationslots L215-01028 to 33 was also assessed under the same conditions. Theresults are shown in Table 42.

TABLE 42 Summary Results for Assay and Related Substances for DrugProducts lots L215-01028 to 33 at T = 0, after 1 month, 3 months and 6months L215-01028 L215-01029 L215-01031 L215-01032 L215-01023 SDIL215-01023 SDI L215-01027 SDI L215-01027 SDI Fenretinide 40%-Fenretinide 40%- Fenretinide 40%- Fenretinide 40%- PVPK30/BHA/BHTPVPK30/BHA/BHT L215-01030 PVPK30/PVPK12/ PVPK30/PVPK12/ L215-01033 SDISDI (L215-01029 core) BHA/BHT SDI BHA/BHT SDI (L215-01032 core)Description Orange capsule Yellow White Orange capsule Yellow Whitefilled with uncoated coated filled with uncoated coated yellow powdertablet tablet yellow powder tablet tablet Storage T = 0 Total deg (%area)  0.47  0.44  0.49  0.47  0.50  0.55 Total Deg in SDI  0.33  0.43(% area) Assay (% LC) 97.7  95.9  90.2  98.9  96.5  84.0  Storage T =month 5° C. Total deg (% area)  0.71  0.72  0.78  0.75  0.76  0.74 TotalDeg in SDI  0.48  0.50 (% area) Assay (% LC) 95.3  95.6  96.4  95.4 94.1  93.0  Storage T = 1 month 25° C./60% RH Total deg (% area)  1.40 1.78  2.02  2.71  2.31  1.24 Total Deg in SDI  0.88  0.81 (% area)Assay (% LC) 92.8  91.4  90.6  87.8  86.5  91.9  Storage T = 3 months 5°C. Total deg (% area)  0.71  0.82  0.86  0.73  0.83  1.02 Total Deg inSDI  1.06  0.74 (% area) Assay (% LC) 96.9  94.4   92.0  94.4  94.9 93.7  Storage T = 3 months 25° C./60% RH Total deg (% area)  6.50  7.11 7.67  8.12  8.51  6.77 Total Deg in SDI  5.68  6.34 (% area) Assay (%LC) 74.2  70.0  65.1  66.8  66.4  72.1  Storage T = 6 months 5° C. Totaldeg (% area)  1.12  1.36  1.92  1.10  1.77  1.93 Total Deg in SDI  2.17 1.65 (% area) Assay (% LC) 91.4  91.4  88.7  91.6  86.7  87.5 

Although the present invention has been described hereinabove by way ofspecific embodiments thereof, it can be modified, without departing fromthe character and nature of the subject invention as defined in theappended claims. The scope of the claims should not be limited by thepreferred embodiments set forth in the examples, but should be given thebroadest interpretation consistent with the description as a whole. Inthe claims, the word “comprising” is used as an open-ended term,substantially equivalent to the phrase “including, but not limited to”.The singular forms “a”, “an” and “the” include corresponding pluralreferences unless the context clearly dictates otherwise.

What is claimed is:
 1. A method for treating or lessening the severityof a disease or condition selected from an inflammatory disease orcondition, a metabolic disease or condition, and cancer, said methodcomprising administering to a subject in need thereof an effectiveamount of an oral dosage formulation comprising an amorphous soliddispersion comprising fenretinide or an analog thereof and at least onematrix polymer selected from a polyvinylpyrrolidone, a hydroxypropylcellulose, a hydroxypropyl methylcellulose hypromellose phthalate, apolyvinylpyrrolidone-vinyl acetate, a hypromellose-acetate-succinate,and any mixture thereof, wherein: (a) at least 55% of said fenretinideor analog thereof in said amorphous solid dispersion is in amorphousform; (b) said fenretinide or analog thereof is present in an amount inthe range of about 20% to about 60% by weight in said amorphous soliddispersion; and (c) said fenretinide analog is4-oxo-N-(4-hydroxyphenyl)retinamide, N-(4-methoxyphenyl)retinamide(4-MPR), 4-hydroxybenzylretinone, 4-(retinamido)phenyl-C-glucuronide,4-(retinamido)phenyl-C-glucoside, 4-(retinamido)benzyl-C-xyloside,1-W-D-glucopyranosyl) retinamide, 1-(D-glucopyranosyluronosyl)retinamide, bexarotene, or a compound of formula I:

wherein R is OH, COOH, CH₂OH, CH₂CH₂OH, or CH₂COOH; carbons a-d and f-iare optionally substituted with one or more groups selected from CH₃,OH, COOH, (CH₃)₂ and CH₂OH, or any combination thereof, and carbon e isoptionally substituted with a C₁-C₃ alkyl group that is optionallysubstituted with CH₃ and/or OH.
 2. The method of claim 1, wherein thematrix polymer comprises a polyvinylpyrrolidone polymer.
 3. The methodof claim 2, wherein the polyvinylpyrrolidone polymer comprisespolyvinylpyrrolidone K30.
 4. The method of claim 1, wherein thefenretinide or analog thereof is present in an amount in the range ofabout 20% to about 30% by weight in said amorphous solid dispersion. 5.The method of claim 1, wherein the [fenretinide or analog thereof/matrixpolymer] ratio in said amorphous solid dispersion is about 1/2 to 2/1.6. The method of claim 1, wherein the amorphous solid dispersioncomprises fenretinide.
 7. The method of claim 1, wherein the formulationcomprises about 50 to about 150 mg of the fenretinide or analog thereof.8. The method of claim 1, wherein the formulation further comprises atleast one pharmaceutical excipient.
 9. The method of claim 8, whereinthe at least one pharmaceutical excipient comprises an antioxidant. 10.The method of claim 9, wherein the antioxidant comprises butylatedhydroxyanisole (BHA), butylated hydroxytoluene (BHT), L-ascorbic acid,or any combination thereof.
 11. The method of claim 8, wherein the atleast one pharmaceutical excipient comprises a disintegrant.
 12. Themethod of claim 11, wherein the disintegrant comprises croscarmellosesodium.
 13. The method of claim 8, wherein the at least onepharmaceutical excipient comprises a filler.
 14. The method of claim 13,wherein the filler comprises microcrystalline cellulose, dibasic calciumphosphate, or a combination thereof.
 15. The method of claim 8, whereinthe at least one pharmaceutical excipient comprises a lubricant.
 16. Themethod of claim 15, wherein the lubricant comprises magnesium stearate.17. The method of claim 1, wherein the oral dosage formulation is in theform of a capsule.
 18. The method of claim 1, wherein the inflammatorydisease or condition comprises inflammation of the respiratory tract.19. The method of claim 1, wherein the inflammatory disease or conditionis cystic fibrosis.
 20. The method of claim 1, wherein the inflammatorydisease or condition comprises neuroinflammation.
 21. The method ofclaim 20, wherein the inflammatory disease or condition is a neuralinjury or a neurodegenerative disease.
 22. The method of claim 21,wherein the inflammatory disease or condition is spinal cord injury,Amyotrophic Lateral Sclerosis, Parkinson's disease, and Huntington'sdisease.
 23. The method of claim 1, wherein the inflammatory disease orcondition is macular degeneration.
 24. The method of claim 1, whereinthe metabolic disease or condition is diabetes or obesity.
 25. Themethod of claim 19, wherein the oral dosage formulation comprises:fenretinide; polyvinylpyrrolidone K30; BHA; BHT; L-ascorbic acid;croscarmellose sodium; microcrystalline cellulose; dibasic calciumphosphate; and magnesium stearate.